Low calorie infant formula with improved physical attributes

ABSTRACT

The present disclosure is directed to low calorie infant formulas, and in particular, low calorie infant formulas that have a low buffering capacity, exhibit an increased rate of protein hydrolysis and digestion, and have an improved tolerance, as compared to full calorie infant formulas. Also disclosed are low calorie liquid infant formulas that have a reduced (i.e., “low”) micronutrient content on a per volume basis, and exhibit an overall improvement in the physical properties of the formula, as compared to low calorie liquid infant formulas having a higher micronutrient content.

CROSS REFERENCE TO RELATED APPLICATIONS

This application claims the benefit of U.S. Provisional Application No.61/428,831 filed Dec. 30, 2010, which disclosure is incorporated byreference in its entirety.

FIELD OF THE DISCLOSURE

The present disclosure is directed to low calorie infant formulas, andin particular, low calorie infant formulas that have a low bufferingcapacity, exhibit an increased rate of protein hydrolysis and digestion,and have improved tolerance, as compared to full calorie infantformulas. Also disclosed are low calorie liquid infant formulas thathave a reduced (i.e., “low”) micronutrient content on a per volumebasis, and exhibit an overall improvement in the physical appearance ofthe formula, including a lighter color and improved stability, ascompared to low calorie liquid infant formulas having a highermicronutrient content.

BACKGROUND OF THE DISCLOSURE

There are numerous types of infant nutritional formulas that are wellknown and widely available. These infant formulas comprise a range ofnutrients designed to meet the nutritional needs of the growing infant,and typically include fats, carbohydrates, proteins, vitamins, minerals,and other nutrients helpful for optimal infant growth and development.

Breast milk, however, is generally recognized as the best nutritionalsource for newborn infants. It is known that human breast milk providesgood immunological benefits to the breastfed infant. Consequently, mostinfant formulas are designed to be closer to breast milk in terms ofcomposition and function.

It is also known that the composition of human breast milk changes overthe first few weeks following delivery of an infant. Human breast milkis referred to as colostrum during the first five days after birth,transition milk during days 6-14 after birth, and mature milkthereafter. During each stage of lactation, the corresponding humanbreast milk composition differs considerably. For instance, colostrumand transition milk have lower caloric densities than mature milk, aswell as higher protein and lower carbohydrate concentrations. Vitaminand mineral concentrations also vary in the three defined human milkgroups.

Some commercial infant formulas are similar in composition, although notidentical, to mature human breast milk, and are used for both newbornsas well as older infants. It has previously been generally accepted thatthe feeding of newborn infants should be conducted with an emphasis onencouraging infant growth, and that such growth is best accomplished byfeeding the infant commercial infant formulas having a similar nutrientand energy content to mature milk.

Recently, attempts have been made to formulate infant formulas fornewborns that have a lower energy content, and thus provide fewercalories during the initial weeks or months of life, than wouldotherwise be provided from feeding with a conventional full calorieinfant formula. Previous attempts at formulating infant formulas havinga low energy content have involved reducing the levels of one or moremacronutrient (e.g., protein, fat, carbohydrate), while maintaining themicronutrient levels at approximately the level found in full calorieinfant formulas on a per volume basis. However, the combination ofreduced macronutrients and high micronutrients can result in a formulawith poor physical attributes. For instance, such formulas are typicallydarker in color, have increased problems with sedimentation, and aremore prone to separation over the shelf life of the product than arefull calorie formulas.

Furthermore, some infant formula fed newborns can experiencegastrointestinal (GI) intolerance problems, including loose stools, gas,and spit-up. The GI intolerance issues may be at least in part due toincomplete nutrient (e.g., protein) digestion and absorption by theinfant. To address this intolerance problem, some infant formulasexclude lactose as an ingredient, while others replace intact milkprotein with hydrolyzed protein to lessen the burden on the infant'sdigestive system.

Some formula fed infants may also experience more episodes of GI tractinfection than do breast fed infants. One explanation for thisphenomenon may be the low buffering capacity of human breast milk. Humanbreast milk is known to have lower acid buffering properties than bothcow milk and cow milk-based infant formulas. The low buffering capacityof human breast milk may allow the natural level of gastric acidity ininfants to be more effective in inactivating orally ingested pathogens.

It would therefore be desirable to provide a low calorie liquid infantformula that has improved physical attributes, such as a lighter colorand improved stability, as compared to previously known low calorieinfant formulas. It would also be desirable to provide an infant formulathat has a low buffering capacity, similar to breast milk, and that alsohas an increased rate of protein hydrolysis and digestion and goodtolerance so as to provide additional benefits to the infant.

SUMMARY OF THE DISCLOSURE

The present disclosure is directed to low calorie liquid infant formulashaving improved physical attributes. These formulas have a reduced(i.e., “low”) micronutrient content on a per volume basis, and exhibitan overall improvement in the physical appearance of the product,including a lighter color and improved stability, as compared to lowcalorie liquid infant formulas having a higher micronutrient content.Also disclosed are low calorie liquid and powder infant formulas thathave a low buffering capacity, exhibit an increased rate of proteinhydrolysis and digestion, and/or have an improved formula tolerance, ascompared to conventional full calorie infant formulas. The low calorieformulas of the present disclosure, when administered to newborn infantsduring the first few weeks of life, provide adequate nutrition for thegrowth and development of the newborn.

Thus, in one embodiment, the present disclosure is directed to a lowmicronutrient infant formula. The formula comprises micronutrients andat least one macronutrient selected from the group consisting ofprotein, carbohydrate, fat, and combinations thereof, and has an energycontent of from about 200 to less than 600 kilocalories per liter offormula. At least 65% of the micronutrients are included in the infantformula in an amount that is from about 30% to about 80% of conventionalamounts of corresponding micronutrients on a per volume basis.

In another embodiment, the present disclosure is directed to a lowmicronutrient infant formula. The formula comprises micronutrients andat least one macronutrient selected from the group consisting ofprotein, carbohydrate, fat, and combinations thereof, and has an energycontent of from about 200 to about 360 kilocalories per liter offormula. At least 45% of the micronutrients are included in the infantformula in an amount that is from about 30% to about 65% of conventionalamounts of corresponding micronutrients, on a per volume basis.

In another embodiment, the present disclosure is directed to a lowmicronutrient infant formula. The formula comprises micronutrients andat least one macronutrient selected from the group consisting ofprotein, carbohydrate, fat, and combinations thereof, and has an energycontent of from about 360 to about 600 kilocalories per liter offormula. At least 30% of the micronutrients are included in the infantformula in an amount that is from about 55% to about 80% of conventionalamounts of corresponding micronutrients, on a per volume basis.

In another embodiment, the present disclosure is directed to a kit. Thekit comprises at least one days 1-2 low micronutrient infant formulacomprising micronutrients and at least one macronutrient selected fromthe group consisting of protein, carbohydrate, fat, and combinationsthereof, and having an energy content of from about 200 to about 360kilocalories per liter of formula, wherein at least 45% of themicronutrients are included in the infant formula in an amount that isfrom about 30% to about 65% of conventional amounts of correspondingmicronutrients, on a per volume basis, and at least one days 3-9 lowmicronutrient infant formula comprising micronutrients and at least onemacronutrient selected from the group consisting of protein,carbohydrate, fat, and combinations thereof, and having an energycontent of from about 360 to less than 600 kilocalories per liter offormula, wherein at least 30% of the micronutrients are included in theinfant formula in an amount that is from about 55% to about 80% ofconventional amounts of corresponding micronutrients, on a per volumebasis.

In another embodiment, the present disclosure is directed to a methodfor providing nutrition to an infant. The method comprises administeringto the infant a low micronutrient infant formula comprisingmicronutrients and at least one macronutrient selected from the groupconsisting of protein, fat, carbohydrates, and combinations thereof, andhaving an energy content of from about 200 to less than 600 kilocaloriesper liter of formula, wherein at least 65% of the micronutrients areincluded in the infant formula in an amount that is from about 30% toabout 80% of conventional amounts of corresponding micronutrients, on aper volume basis.

In another embodiment, the present disclosure is directed to a methodfor providing nutrition to an infant. The method comprises administeringto the infant a low micronutrient infant formula comprisingmicronutrients and at least one macronutrient selected from the groupconsisting of protein, carbohydrate, fat, and combinations thereof, andhaving an energy content of from about 200 to about 360 kilocalories perliter of formula, wherein at least 45% of the micronutrients areincluded in the infant formula in an amount that is from about 30% toabout 65% of conventional amounts of corresponding micronutrients, on aper volume basis.

It has now surprisingly been discovered that low calorie liquid infantformulas having improved physical attributes can be formulated if asufficient amount of one or more micronutrients in the low calorieformula is generally matched to that of full calorie formulas on a perkilocalorie (kcal) basis, rather than on a per volume basis. Theseformulas thus have a reduced (i.e., “low”) micronutrient content on aper volume basis, and exhibit an overall improvement in the physicalappearance of the product, including a lighter color and improvedstability, than do low calorie liquid infant formulas having a highermicronutrient content.

It has also been discovered that the low calorie liquid or powder infantformulas have a lower buffering capacity than conventional full calorieinfant formulas, and in some embodiments, have a lower bufferingcapacity than that of human milk. The low calorie infant formulas of thepresent disclosure can thus be used to regulate gastric acidity ininfants, reduce the growth of pathogenic microorganisms in the infant GItract, and promote the growth of beneficial microorganisms. The lowcalorie infant formulas of the present disclosure have also been foundto exhibit an increased rate of protein hydrolysis and digestion, andthus have an improved formula tolerance, as compared to conventional,full calorie infant formulas.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is a chart showing the buffering strength of various low caloriedays 1-2 and days 3-9 infant formulas, as compared to control fullcalorie formulas and to human milk, as discussed in Example 16.

FIG. 2 is a chart showing the buffering capacity of various low caloriedays 1-2 and days 3-9 infant formulas, as compared to control fullcalorie formulas and to human milk, as discussed in Example 16.

FIG. 3 is a chart showing the effect of HCl addition on the pH of lowcalorie days 1-2 and days 3-9 reconstituted powder infant formulas, ascompared to a control full calorie formula, as discussed in Example 17.

FIG. 4 is a chart showing the buffering strength of low calorie days 1-2and days 3-9 reconstituted powder infant formulas, as compared to acontrol full calorie formula, as discussed in Example 17.

FIG. 5 is a chart showing the buffering capacity, as measured by pHdecrease following addition of 5.50 mmoles of HCl to 100 mL of formula,of low calorie days 1-2 and days 3-9 reconstituted powder infantformulas, as compared to a control full calorie formula, as discussed inExample 17.

FIG. 6 is a chart showing the buffering capacity, as measured byincrease in [H+] following addition of 5.50 mmoles of HCl to 100 mL offormula, of low calorie days 1-2 and days 3-9 reconstituted powderinfant formulas, as compared to a control full calorie formula, asdiscussed in Example 17.

FIG. 7 is a chart showing the protein molecular weight (MW) median forlow calorie days 1-2 and days 3-9 reconstituted powder infant formulasfollowing in vitro gastrointestinal digestion, as compared to a controlfull calorie formula, as discussed in Example 20.

FIG. 8 is a chart showing the percent total protein having a MW greaterthan 5000 Da for low calorie days 1-2 and days 3-9 reconstituted powderinfant formulas following in vitro gastrointestinal digestion, ascompared to a control full calorie formula, as discussed in Example 20.

FIG. 9 is a chart showing the amount of insoluble (indigestible) proteinin the protein pellet following high speed centrifugation of low caloriedays 1-2 and days 3-9 reconstituted powder infant formulas following invitro gastrointestinal digestion, as compared to a control full calorieformula, as discussed in Example 20.

FIG. 10 is a chart showing the protein MW median for low calorie days1-2 and days 3-9 reconstituted powder infant formulas followingpancreatin digestion for 71 minutes, as compared to a control fullcalorie formula, as discussed in Example 23.

FIG. 11 is a chart showing the percent total protein having a MW greaterthan 5000 Da for low calorie days 1-2 and days 3-9 reconstituted powderinfant formulas following pancreatin digestion for 71 minutes, ascompared to a control full calorie formula, as discussed in Example 23.

FIG. 12 is a chart showing the particle size distribution for retortsterilized days 1-2 formulas having either a high micronutrient content(Formula 3) or a low micronutrient content (Formula 1), as discussed inExample 29.

DETAILED DESCRIPTION OF THE DISCLOSURE

The low calorie liquid infant formulas disclosed herein may have a lowmicronutrient content, on a per volume basis, and improved physicalattributes as compared to conventional infant formulas that have ahigher micronutrient content. Further, the methods of the presentdisclosure utilize low calorie liquid and powder infant formulas toregulate gastric acidity in infants, reduce the growth of pathogenicmicroorganisms and promote the growth of beneficial microorganisms inthe infant GI tract, increase the rate of protein hydrolysis anddigestion, and improve formula tolerance. These and other optionalfeatures of the infant formulas and methods of the present disclosure,as well as some of the many other optional variations and additions, aredescribed in detail hereafter.

The terms “retort” and “retort sterilized” are used interchangeablyherein, and unless otherwise specified, refer to the common practice offilling a container, most typically a metal can or other similarpackage, with a nutritional liquid, such as a liquid infant formula, andthen subjecting the liquid-filled package to the necessary heatsterilization step, to form a retort sterilized nutritional liquidproduct.

The terms “aseptic” and “aseptic sterilized” are used interchangeablyherein, and unless otherwise specified, refer to the manufacture of apackaged product without reliance upon the above-described retortpackaging step, wherein the nutritional liquid and package aresterilized separately prior to filling, and then are combined understerilized or aseptic processing conditions to form a sterilized,aseptically packaged, nutritional liquid product.

The terms “nutritional formula” or “nutritional product” or “nutritionalcomposition,” as used herein, are used interchangeably and, unlessotherwise specified, refer to nutritional liquids, nutritionalsemi-liquids, nutritional solids, nutritional semi-solids, nutritionalpowders, nutritional supplements, and any other nutritional food productas known in the art. The nutritional solids and powders may bereconstituted to form a nutritional liquid, all of which comprise one ormore of fat, protein and carbohydrate, and are suitable for oralconsumption by a human. Nutritional formulas may include infantformulas.

The term “nutritional liquid,” as used herein, unless otherwisespecified, refers to nutritional products in ready-to-drink liquid form,concentrated form, and nutritional liquids made by reconstituting thenutritional powders described herein prior to use.

The term “nutritional powder,” as used herein, unless otherwisespecified, refers to nutritional products in flowable or scoopable formthat can be reconstituted with water or another aqueous liquid prior toconsumption and includes both spray dried and drymixed/dryblendedpowders.

The term “nutritional semi-liquid,” as used herein, unless otherwisespecified, refers to those forms that are intermediate in properties,such as flow properties, between liquids and solids, examples of whichinclude thick shakes and liquid gels.

The term “nutritional semi-solid,” as used herein, unless otherwisespecified, refers to those forms that are intermediate in properties,such as rigidity, between solids and liquids, examples of which includepuddings, gelatins, and doughs.

The term “infant,” as used herein, unless otherwise specified, refers toa child 12 months or younger. The term “preterm infant,” as used herein,refers to an infant born prior to 36 weeks of gestation. The term “terminfant,” as used herein, refers to an infant born at or after 36 weeksof gestation.

The term “newborn infant,” as used herein, unless otherwise specified,refers to infants less than about 3 months of age, including infantsfrom zero to about 2 weeks of age. The newborn infant may be a term orpreterm infant.

The term “infant formula,” as used herein, unless otherwise specified,refers to liquid and solid nutritional products suitable for consumptionby an infant. Unless otherwise specified herein, the term “infantformula” is intended to encompass both term and preterm infant formulas.

The term “preterm infant formula,” as used herein, unless otherwisespecified, refers to liquid and solid nutritional products suitable forconsumption by a preterm infant.

The term “micronutrient,” as used herein, refers to essential substancesneeded by organisms in small quantities. Non-limiting examples includevitamins, minerals, and the like.

The term “full calorie infant formula,” as used herein, refers to aninfant formula in which the caloric density or energy content of theformula has not been reduced from that conventionally included in infantformula. Typically, a full calorie infant formula will have an energycontent of at least 600 kcal/L, or even at least 660 kcal/L, and moretypically at least 676 kcal/L, including 600 kcal/L to 800 kcal/L.

The term “low calorie infant formula,” as used herein, refers to aninfant formula that has a lower energy content, on a per volume basis,than a full calorie infant formula.

The terms “high micronutrient” or “high micronutrient content,” whenreferring to the micronutrient content of an infant formula, means atleast 80% of the micronutrients in the infant formula are present inamounts approximately the same as (typically within about 82% for mostmicronutrients) the amount of the micronutrients conventionally includedin infant formulas.

All percentages, parts and ratios as used herein, are by weight of thetotal composition, unless otherwise specified. All such weights, as theypertain to listed ingredients, are based on the active level and,therefore, do not include solvents or by-products that may be includedin commercially available materials, unless otherwise specified.

Numerical ranges as used herein are intended to include every number andsubset of numbers within that range, whether specifically disclosed ornot. Further, these numerical ranges should be construed as providingsupport for a claim directed to any number or subset of numbers in thatrange. For example, a disclosure of from 1 to 10 should be construed assupporting a range of from 2 to 8, from 3 to 7, from 5 to 6, from 1 to9, from 3.6 to 4.6, from 3.5 to 9.9, and so forth.

All references to singular characteristics or limitations of the presentdisclosure shall include the corresponding plural characteristic orlimitation, and vice versa, unless otherwise specified or clearlyimplied to the contrary by the context in which the reference is made.

All combinations of method or process steps as used herein can beperformed in any order, unless otherwise specified or clearly implied tothe contrary by the context in which the referenced combination is made.

The various embodiments of the infant formulas and methods of thepresent disclosure may also be substantially free of any optional orselected ingredient or feature described herein, provided that theremaining infant formulas still contains all of the required ingredientsor features as described herein. In this context, and unless otherwisespecified, the term “substantially free” means that the selected infantformulas contains less than a functional amount of the optionalingredient, typically less than 1%, including less than 0.5%, includingless than 0.1%, and also including zero percent, by weight of suchoptional or selected ingredient.

The infant formulas and methods of the present disclosure may comprise,consist of, or consist essentially of the elements of the products andmethods as described herein, as well as any additional or optionalelement described herein or otherwise useful in nutritional infantformula applications.

Product Form

The infant formulas of the present disclosure may be formulated andadministered in any known or otherwise suitable oral product form. Anysolid, semi-solid, liquid, semi-liquid, or powder form, includingcombinations or variations thereof, are suitable for use herein,provided that such forms allow for safe and effective oral delivery tothe individual of the essential ingredients as also defined herein.

Specific non-limiting examples of product forms suitable for use withproducts and methods disclosed herein include, for example, liquid andpowder preterm infant formulas, liquid and powder term infant formulas,and liquid and powder elemental and semi-elemental formulas.

The infant formulas of the present disclosure are preferably formulatedas dietary product forms, which are defined herein as those embodimentscomprising the essential ingredients of the present disclosure in aproduct form that then contains at least one of fat, protein, andcarbohydrate.

The infant formulas may be formulated with sufficient kinds and amountsof nutrients to provide a sole, primary, or supplemental source ofnutrition, or to provide a specialized nutritional product for use ininfants afflicted with specific diseases or conditions or with atargeted nutritional benefit.

Desirably, the infant formulas of the present disclosure are formulatedfor newborn infants, including both term and preterm newborn infants.Preferably, the infant formula is formulated for feeding to newborninfants within the first few weeks following birth, and more preferablyfor feeding to newborn infants from age zero to two weeks. In oneembodiment, the infant formula is formulated for feeding to newborninfants in the first two days following birth. Such a formula isreferred to herein as a “days 1-2 formula” or a “days 1-2 infantformula.” In other embodiments, the infant formula is formulated forfeeding to newborn infants during days 3-9 following birth. Such aformula is referred to herein a “days 3-9 formula” or a “days 3-9 infantformula.” It is to be understood that the administration of a days 1-2infant formula of the present disclosure is not limited toadministration during only the first two days following birth, but maybe administered to older infants as well in some embodiments. Similarly,the administration of a days 3-9 infant formula is not limited toadministration during only days 3-9 following birth, but may beadministered to infants of other ages as well in some embodiments.

Nutritional Liquids

Nutritional liquids include both concentrated and ready-to-feednutritional liquids. These nutritional liquids are most typicallyformulated as suspensions, emulsions or clear or substantially clearliquids.

Nutritional emulsions suitable for use may be aqueous emulsionscomprising proteins, fats, and carbohydrates. These emulsions aregenerally flowable or drinkable liquids at from about 1° C. to about 25°C. and are typically in the form of oil-in-water, water-in-oil, orcomplex aqueous emulsions, although such emulsions are most typically inthe form of oil-in-water emulsions having a continuous aqueous phase anda discontinuous oil phase.

The nutritional liquids may be and typically are shelf stable. Thenutritional liquids typically contain up to about 95% by weight ofwater, including from about 50% to about 95%, also including from about60% to about 90%, and also including from about 70% to about 85%, ofwater by weight of the nutritional liquid. The nutritional liquids mayhave a variety of product densities, but most typically have a densitygreater than about 1.03 g/mL, including greater than about 1.04 g/mL,including greater than about 1.055 g/mL, including from about 1.06 g/mLto about 1.12 g/mL, and also including from about 1.085 g/mL to about1.10 g/mL.

The nutritional liquid may have a pH ranging from about 3.5 to about 8,but are most advantageously in a range of from about 4.5 to about 7.5,including from about 5.5 to about 7.3, including from about 6.2 to about7.2.

Although the serving size for the nutritional liquid can vary dependingupon a number of variables, a typical serving size is generally at leastabout 2 mL, or even at least about 5 mL, or even at least about 10 mL,or even at least about 25 mL, including ranges from about 2 mL to about300 mL, including from about 100 mL to about 300 mL, from about 4 mL toabout 250 mL, from about 150 mL to about 250 mL, from about 10 mL toabout 240 mL, and from about 190 mL to about 240 mL.

Nutritional Powders

The nutritional powders are in the form of flowable or substantiallyflowable particulate compositions, or at least particulate compositions.Particularly suitable nutritional powder forms include spray dried,agglomerated or dryblended powder compositions, or combinations thereof,or powders prepared by other suitable methods. The compositions caneasily be scooped and measured with a spoon or similar other device,wherein the compositions can easily be reconstituted with a suitableaqueous liquid, typically water, to form a nutritional liquid, such asan infant formula, for immediate oral or enteral use. In this context,“immediate” use generally means within about 48 hours, most typicallywithin about 24 hours, preferably right after or within 20 minutes ofreconstitution.

Energy Content

The infant formulas of the present disclosure have low energy content(used herein interchangeably with the term “caloric density”) relativeto conventional term and preterm infant formulas. Specifically, theinfant formulas of the present disclosure provide a caloric density orenergy content of from about 200 kcal/L to less than 600 kcal/L,including from about 200 kcal/L to about 500 kcal/L, and moreparticularly from about 250 kcal/L to about 500 kcal/L. The days 1-2infant formulas of the present disclosure provide a caloric density orenergy content of from about 200 kcal/L to about 360 kcal/L, includingfrom about 200 kcal/L to about 350 kcal/L, also including from about 250kcal/L to about 350 kcal/L, from about 250 kcal/L to about 310 kcal/L,and more particularly about 250 kcal/L or about 270 kcal/L. The days 3-9infant formulas of the present disclosure provide a caloric density orenergy content of from about 360 kcal/L to less than 600 kcal/L,including from about 370 kcal/L to less than 600 kcal/L, also includingfrom about 360 kcal/L to about 500 kcal/L, from about 390 kcal/L toabout 470 kcal/L, and in particular about 406 kcal/L or about 410kcal/L. In contrast to the infant formulas of the present disclosure,the caloric density or energy content of conventional term and preterminfant formulas, which are also referred to herein as “full calorieinfant formulas,” is significantly higher, typically ranging from 600kcal/L to 880 kcal/L.

When the infant formulas of the present disclosure are in powder form,then the powder is intended for reconstitution prior to use to obtainthe above-noted caloric densities and other nutrient requirements asdescribed herein. Likewise, when the infant formulas of the presentdisclosure are in a concentrated liquid form, then the concentrate isintended for dilution prior to use to obtain the requisite caloricdensities and nutrient requirements. The infant formulas can also beformulated as ready-to-feed liquids already having the requisite caloricdensities and nutrient requirements.

The infant formulas of the present disclosure are desirably administeredto infants, and in particular newborn infants, in accordance with themethods described in detail herein. Such methods may include feedingswith the infant formulas in accordance with the daily formula intakevolumes described herein.

The energy component of the infant formula is most typically provided bya combination of fat, protein, and carbohydrate nutrients. The proteinmay comprise from about 4% to about 40% of the total calories, includingfrom about 10% to about 30%, also including from about 15% to about 25%;the carbohydrate may comprise less than 40% of the total calories,including from about 5% to about 37%, also including less than about36%, and also including from about 20% to about 33%; and the fat maycomprise the remainder of the formula calories, most typically less thanabout 60% of the calories, including from about 30% to about 60%. Otherexemplary amounts are set forth hereinafter.

Micronutrients

In addition to a low energy content, in some embodiments, the infantformulas of the present disclosure are also characterized by a lowmicronutrient content, on a per volume basis.

As described herein, previous attempts at formulating infant formulashaving a low energy content have involved reducing the levels of one ormore macronutrients (e.g., protein, fat, carbohydrate), whilemaintaining the micronutrient level at approximately the level found infull calorie infant formulas on a per volume basis. For example, oneliter of such a low calorie formula would have reduced amounts of one ormore macronutrient, as compared to one liter of the full calorieformula, but approximately the same amount (typically within at leastabout 82% for most micronutrients) of micronutrients as are found in oneliter of the full calorie formula. However, the combination of reducedmacronutrients and high micronutrients results in a formula with poorphysical attributes. For instance, such formulas are typically darker incolor, have increased problems with sedimentation, and are more prone toseparation over the shelf life of the product than are full calorieformulas.

It has now surprisingly been discovered that low calorie liquid infantformulas having improved physical attributes can be formulated if theamount of micronutrients in the low calorie formula is generally matchedto that of full calorie formulas on a per kilocalorie (kcal) basis,rather than on a per volume basis. For example, 100 kcal of the lowcalorie formula would comprise approximately the same amount (typicallywithin about 80% for most micronutrients) of micronutrients as are foundin 100 kcal of the full calorie formula. In this example, themicronutrient content of the low calorie formula would be formulated ona 100 kcal basis. Low calorie liquid infant formulas that are formulatedon a per kcal basis have a reduced (i.e., “low”) micronutrient contenton a per volume basis (i.e., as compared to the same volume of a fullcalorie formula), and exhibit an overall improvement in the physicalappearance of the formula, including a lighter color and improvedstability.

Thus, in some embodiments, the present disclosure is directed to lowcalorie, low micronutrient infant formulas. As used herein, the term“low micronutrient” or “low micronutrient content,” when referring toinfant formula, means the amount of at least a portion of themicronutrients included in the infant formula is lower than the amountof the corresponding micronutrients conventionally included in infantformula, on a per volume basis. It should be understood that it is notnecessary for the amount of all micronutrients included in an infantformula to be lower than the conventional amounts of correspondingmicronutrients, on a per volume basis, in order for the infant formulato be considered a low micronutrient infant formula. Reduction of aportion of the micronutrients in the infant formula, as compared toconventional amounts on a per volume basis, is sufficient.

The amount of micronutrients “conventionally included in infant formula”or “conventional amounts” of micronutrients refers to industryrecognized standard amounts of micronutrients, on a per volume basis,for inclusion in infant formula in order to achieve adequate growth anddevelopment of infants. Conventional amounts of select micronutrientsthat may be included in infant formula, on a per volume basis, are setforth in Table A (ready-to-feed formulas) and Table B (reconstitutedpowder formulas) below.

TABLE A Ready-to-Feed Formulas Typical Typical amount amount for a foran Minimum Maximum retort aseptic amount amount formula formulaMicronutrient (per L) (per L) (per L) (per L) Vitamin A (IU) 2030 44003110 3890 Vitamin D (IU) 406 642 526 506 Vitamin E (IU) 10.2 15.0 13.311.8 Vitamin K (μg) 54.1 410 125 106 Thiamin (μg) 676 4060 1220 1420Riboflavin (μg) 1010 4000 2500 2590 Vitamin B6 (μg) 406 556 476 495Vitamin B12 (μg) 1.69 14.0 4.7 5.4 Niacin (μg) 7100 21000 9730 9680Folic acid (μg) 101 600 193 212 Pantothenic acid (μg) 3040 14400 62206710 Biotin (μg) 29.7 169 56.1 67.2 Vitamin C (mg) 60.8 800 416 352Choline (mg) 109 203 127 120 Inositol (mg) 31.8 130 39.8 39.9 Calcium(mg) 528 620 585 581 Phosphorus (mg) 284 398 349 341 Magnesium (mg) 40.671.5 55.7 55.0 Iron (mg) 12.2 15.6 13.4 13.7 Zinc (mg) 5.07 14.0 6.466.67 Manganese (μg) 33.8 235 84.4 87.8 Copper (μg) 609 1484 676 728Iodine (μg) 40.2 474 118 140 Sodium (mg) 163 245 190 189 Potassium (mg)710 1196 946 942 Chloride (mg) 440 551 474 504 Fluoride (μg) — — 168 143Selenium (μg) 12.3 36.1 24.9 24.3

TABLE B Reconstituted Powder Formulas Minimum Maximum Typical amountamount amount Micronutrient (per L) (per L) (per L) Vitamin A (IU) 20304820 3583 Vitamin D (IU) 406 642 563 Vitamin E (IU) 10.1 15.0 12.6Vitamin K (μg) 54.1 410 137 Thiamin (μg) 676 4060 1560 Riboflavin (μg)1010 4000 1500 Vitamin B6 (μg) 406 556 467 Vitamin B12 (μg) 1.69 14.05.85 Niacin (μg) 7100 21000 9400 Folic acid (μg) 101 600 209 Pantothenicacid (μg) 3040 14400 6750 Biotin (μg) 29.7 169 63.8 Vitamin C (mg) 60.8670 170 Choline (mg) 108 203 123 Inositol (mg) 31.8 130 41.0 Calcium(mg) 536 637 580 Phosphorus (mg) 289 408 332 Magnesium (mg) 40.6 73.353.7 Iron (mg) 12.4 16.1 13.9 Zinc (mg) 5.15 14.4 6.69 Manganese (μg)34.3 148 89.7 Copper (μg) 618 1519 720 Iodine (μg) 41.0 489 126 Sodium(mg) 165 251 201 Potassium (mg) 721 1235 1039 Chloride (mg) 446 565 486Fluoride (μg) — — 116 Selenium (μg) 12.4 37.0 25.6

Exemplary non-limiting micronutrients that may be included inconventional infant formulas include vitamin A, vitamin D, vitamin E,vitamin K, thiamin, riboflavin, vitamin B6, vitamin B12, niacin, folicacid, pantothenic acid, biotin, vitamin C, choline, inositol, calcium,phosphorus, magnesium iron, zinc, manganese, copper, iodine, sodium,potassium, chloride, fluoride, selenium, and combinations thereof. Someexemplary conventional infant formula may include a combination ofcopper, phosphorus, iron, calcium, and zinc. Some other exemplaryconventional infant formulas may include a combination of copper, ironand phosphorus.

In one specific embodiment, at least two of copper, phosphorus, iron,calcium, and zinc are present in the low micronutrient formulas inamount of about 5% less, or even 10% less, or even 20% less, or even 30%less, or even 50% less, or even 75% less, or even 80% less, or even 90%less than the amounts set forth in Tables A and B above. In anotherspecific embodiment, iron and copper are present in the lowmicronutrient formulas in amount of about 5% less, or even 10% less, oreven 20% less, or even 30% less, or even 50% less, or even 75% less, oreven 80% less, or even 90% less than the amounts set forth in Tables Aand B above.

It should be understood that Tables A and B do not contain an exhaustivelist of suitable micronutrients that can be included in the infantformulas of the present disclosure. Further, the low micronutrientinfant formulas of the present disclosure need not comprise everymicronutrient listed in Tables A and B. The present disclosurecontemplates infant formulas comprising any combination of one or moreof the micronutrients listed in Tables A and B and/or othermicronutrients known in the art as suitable for inclusion in infantformula. Standard or conventional amounts of these and othermicronutrients (on a per 100 kcal basis) can readily be determined withreference to European and/or United States infant formula regulationsand standards.

When determining whether the micronutrient content in an infant formulais low, on a per volume basis, as compared to conventional amounts, theamounts of “corresponding micronutrients” should be compared. In thisinstance, “corresponding micronutrients” refers to the samemicronutrients as are present in the infant formula being evaluated. Forexample, if the infant formula comprises the micronutrients calcium,phosphorus, and magnesium, the amounts of these micronutrients in theinfant formula should be compared to the amounts of calcium, phosphorus,and magnesium, respectively, that are conventionally included in infantformula, to determine if the amount of these micronutrients in theinfant formula is “low.”

The amount of micronutrients included in the low micronutrient infantformulas of the present disclosure can be expressed as a percentage ofthe conventional amounts of corresponding micronutrients, on a pervolume basis. For instance, in some embodiments of the presentdisclosure, low micronutrient infant formulas are provided wherein themicronutrients are included in the infant formula in an amount that isfrom about 30% to about 80% of conventional amounts of correspondingmicronutrients, on a per volume basis, including from about 30% to about65%, from about 55% to about 80%, from about 40% to about 70%, fromabout 40% to about 50%, and from about 60% to about 70% of conventionalamounts of corresponding micronutrients, all on a per volume basis.Typically, at least 65% of the micronutrients, including at least 75%,at least 80%, at least 90%, and 100% of the micronutrients in the lowmicronutrient infant formulas of the present disclosure are included inthe infant formula in amounts that are from about 30% to about 80% ofconventional amounts of corresponding micronutrients, on a per volumebasis.

In some embodiments, low micronutrient infant formulas are providedwherein the micronutrients are included in the infant formula in anamount that is from about 30% to about 65% of conventional amounts ofcorresponding micronutrients, on a per volume basis, including fromabout 35% to about 60%, from about 40% to about 50%, from about 40% toabout 45%, and in particular about 40% of conventional amounts ofcorresponding micronutrients, all on a per volume basis. In suchembodiments, typically at least 45% of the micronutrients, including atleast 50%, at least 60% at least 75%, at least 80%, at least 90%, and100% of the micronutrients in the low micronutrient infant formula areincluded in the infant formula in amounts that are from about 35% toabout 60% of conventional amounts of corresponding micronutrients, on aper volume basis. In other embodiments, at least 10% of themicronutrients, including at least 25%, at least 50%, at least 60%, atleast 75%, and at least 80% of the micronutrients in the lowmicronutrient infant formula are included in the infant formula inamounts that are from about 40% to about 50% of conventional amounts ofcorresponding micronutrients, on a per volume basis. Such lowmicronutrient infant formulas may include, for example, days 1-2 infantformulas.

In other embodiments, low micronutrient infant formulas are providedwherein the micronutrients are included in the infant formula in anamount that is from about 55% to about 80% of conventional amounts ofcorresponding micronutrients, on a per volume basis, including fromabout 60% to about 75%, from about 60% to about 70%, from about 60% toabout 65%, and in particular about 60% of conventional amounts ofcorresponding micronutrients, all on a per volume basis. In suchembodiments, typically at least 30% of the micronutrients, including atleast 50%, at least 60%, at least 75%, at least 80%, at least 90%, and100% of the micronutrients in the low micronutrient infant formula areincluded in the infant formula in amounts that are from about 55% toabout 80% of conventional amounts of corresponding micronutrients, on aper volume basis. In other embodiments, at least 10%, including at least25%, at least 50%, at least 60%, at least 75%, and at least 80%, of themicronutrients in the low micronutrient infant formula are included inthe infant formula in amounts that are from about 60% to about 70% ofconventional amounts of corresponding micronutrients, on a per volumebasis. Such low micronutrient infant formulas may include, for example,days 3-9 infant formulas.

In some embodiments where the micronutrient includes minerals, theminerals are included in the low micronutrient infant formula in anamount that is from about 30% to about 80% of conventional amounts ofcorresponding minerals, on a per volume basis, including from about 30%to about 65%, from about 55% to about 80%, from about 40% to about 70%,from about 40% to about 50%, and from about 60% to about 70% ofconventional amounts of corresponding minerals, all on a per volumebasis. Typically, at least 10%, including at least 45%, at least 50%, atleast 60%, at least 70%, at least 75%, at least 80%, at least 90%, and100%, of the minerals in the low micronutrient infant formulas of thepresent disclosure are included in the infant formula in amounts thatare from about 30% to about 80% of conventional amounts of correspondingminerals, on a per volume basis.

In still other embodiments, the minerals are included in the lowmicronutrient infant formula in an amount that is from about 30% toabout 65% of conventional amounts of corresponding minerals, on a pervolume basis, including from about 35% to about 60%, from about 40% toabout 50%, from about 40% to about 45%, and in particular about 40% ofconventional amounts of corresponding minerals, all on a per volumebasis. In such embodiments, typically at least 10%, including at least25%, at least 50%, at least 60%, at least 75%, at least 80%, at least90%, and 100%, of the minerals in the low micronutrient infant formulaare included in the infant formula in amounts that are from about 30% toabout 65% of conventional amounts of corresponding minerals, on a pervolume basis. In other embodiments, at least 10%, including at least25%, at least 50%, at least 60%, at least 75%, at least 80%, at least90%, and 100%, of the minerals in the low micronutrient infant formulaare included in the infant formula in amounts that are from about 40% toabout 50% of conventional amounts of corresponding minerals, on a pervolume basis. Such low micronutrient infant formulas may include, forexample, days 1-2 infant formulas.

In still other embodiments, the minerals are included in the lowmicronutrient infant formula in an amount that is from about 55% toabout 80% of conventional amounts of corresponding minerals, on a pervolume basis, including from about 60% to about 75%, from about 60% toabout 70%, from about 60% to about 65%, and in particular about 60% ofconventional amounts of corresponding minerals, all on a per volumebasis. In such embodiments, typically at least 10%, including at least25%, at least 50%, at least 60%, at least 75%, at least 80%, at least90%, and 100%, of the minerals in the low micronutrient infant formulaare included in the infant formula in amounts that are from about 55% toabout 80% of conventional amounts of corresponding minerals, on a pervolume basis. In other embodiments, at least 10%, including at least25%, at least 50%, at least 60%, at least 75%, at least 80%, at least90%, and 100%, of the minerals in the low micronutrient infant formulaare included in the infant formula in amounts that are from about 60% toabout 70% of conventional amounts of corresponding minerals, on a pervolume basis. Such low micronutrient infant formulas may include, forexample, days 3-9 infant formulas.

In some embodiments where the micronutrient includes vitamins, thevitamins are included in the low micronutrient infant formula in anamount that is from about 30% to about 80% of conventional amounts ofcorresponding vitamins, on a per volume basis, including from about 30%to about 65%, from about 55% to about 80%, from about 40% to about 70%,from about 40% to about 50%, and from about 60% to about 70% ofconventional amounts of corresponding vitamins, all on a per volumebasis. Typically, at least 45%, including at least 50%, at least 60%, atleast 70%, at least 80%, at least 85%, at least 90%, and 100%, of thevitamins in the low micronutrient infant formulas of the presentdisclosure are included in the infant formula in amounts that are fromabout 30% to about 80% of conventional amounts of correspondingvitamins, on a per volume basis.

In still other embodiments, the vitamins are included in the lowmicronutrient infant formula in an amount that is from about 30% toabout 65% of conventional amounts of corresponding vitamins, on a pervolume basis, including from about 35% to about 60%, from about 40% toabout 50%, from about 40% to about 45%, and in particular about 40% ofconventional amounts of corresponding vitamins, all on a per volumebasis. In such embodiments, typically at least 10%, including at least25%, at least 50%, at least 60%, at least 75%, at least 80%, at least90%, and 100%, of the vitamins in the low micronutrient infant formulaare included in the infant formula in amounts that are from about 30% toabout 65% of conventional amounts of corresponding vitamins, on a pervolume basis. In other embodiments, at least 10%, including at least25%, at least 50%, at least 60%, at least 75%, and at least 80%, of thevitamins in the low micronutrient infant formula are included in theinfant formula in amounts that are from about 40% to about 50% ofconventional amounts of corresponding vitamins, on a per volume basis.Such low micronutrient infant formulas may include, for example, days1-2 infant formulas.

In still other embodiments, the vitamins are included in the lowmicronutrient infant formula in an amount that is from about 55% toabout 80% of conventional amounts of corresponding vitamins, on a pervolume basis, including from about 60% to about 75%, from about 60% toabout 70%, from about 60% to about 65%, and in particular about 60% ofconventional amounts of corresponding vitamins, all on a per volumebasis. In such embodiments, typically at least 10%, including at least25%, at least 50%, at least 60%, at least 75%, at least 80%, at least90%, and 100%, of the vitamins in the low micronutrient infant formulaare included in the infant formula in amounts that are from about 55% toabout 80% of conventional amounts of corresponding vitamins, on a pervolume basis. In other embodiments, at least 10%, including at least25%, at least 50%, at least 60%, at least 75%, at least 80%, and atleast 90%, of the vitamins in the low micronutrient infant formula areincluded in the infant formula in amounts that are from about 60% toabout 70% of conventional amounts of corresponding vitamins, on a pervolume basis. Such low micronutrient infant formulas may include, forexample, days 3-9 infant formulas.

Suitable micronutrients for inclusion in the infant formulas of thepresent disclosure include vitamins or related nutrients, minerals, andcombinations thereof. Non-limiting examples of suitable vitamins includevitamin A, vitamin D, vitamin E, vitamin K, thiamine, riboflavin,pyridoxine, vitamin B5, vitamin B6, vitamin B12, niacin, folic acid,pantothenic acid, biotin, vitamin C, choline, inositol, ascorbic acid,salts and derivatives thereof, and combinations thereof.

Non-limiting examples of suitable minerals that may be included in theinfant formulas of the present disclosure include calcium, phosphorus,magnesium, iron, zinc, manganese, copper, iodine, sodium, potassium,molybdenum, chromium, chloride, fluoride, selenium, and combinationsthereof.

Any infant formula may be formulated with a low micronutrient content asdisclosed herein, including both retort and aseptic ready-to-feednutritional liquids, concentrated nutritional liquids, and nutritionalpowders.

Macronutrients

The infant formulas of the present disclosure may further comprise oneor more macronutrient, in addition to the micronutrients describedherein. The macronutrients include protein, fat, carbohydrate, andcombinations thereof. Macronutrients suitable for use herein include anyprotein, fat, carbohydrate, or source thereof that is known for orotherwise suitable for use in an oral nutritional product, provided thatthe macronutrient is safe and effective for oral administration toinfants and is otherwise compatible with the other ingredients in theinfant formula.

Although total concentrations or amounts of the protein, fat, andcarbohydrate may vary depending upon the product form (e.g., powder orready-to-feed liquid) and targeted dietary needs of the intended user,such concentrations or amounts most typically fall within one of theembodied ranges described in the following table (each numerical valueis preceded by the term “about”), inclusive of any other essential fat,protein, and/or carbohydrate ingredients as described herein. For powderembodiments, the amounts in the following table are amounts followingreconstitution of the powder.

TABLE C Nutrient (g/100 mL) Example A Example B Protein 0.5 to 1.0 0.6to 0.9 Fat 1.2 to 2.5 1.4 to 2.3 Carbohydrate 2.7 to 6.5 3.1 to 6.1

The total concentrations or amounts of the protein, fat, andcarbohydrate may also vary depending upon whether the infant formula isa days 1-2 formula or a days 3-9 formula. The concentration of protein,fat, and carbohydrate for the days 1-2 and the days 3-9 formulas aremost typically formulated within any of the embodied ranges described inthe following table (each numerical value is preceded by the term“about”), inclusive of any other essential fat, protein, and/orcarbohydrate ingredients as described herein. For powder embodiments,the amounts in the following table are amounts following reconstitution.

TABLE D Days 1-2 Formula Days 3-9 Formula Nutrient (g/100 mL) Example CExample D Example E Example F Protein 0.50 to 0.75 0.58 to 0.72 0.76 to1.0  0.85 to 0.98 Fat 1.2 to 1.7 1.4 to 1.6 1.8 to 2.5 2.0 to 2.2Carbohydrate 2.7 to 4.0 2.9 to 3.6 4.1 to 6.5 4.9 to 6.3

The level or amount of carbohydrate, fat, and protein in the infantformula (whether a powder formula or a liquid ready-to-feed orconcentrated liquid) may also be characterized in addition to or in thealternative as a percentage of total calories in the infant formulas.These macronutrients for infant formulas of the present disclosure aremost typically formulated within any of the caloric ranges described inthe following table (each numerical value is preceded by the term“about”).

TABLE E Nutrient (% total calories) Example G Example H Example ICarbohydrate 2 to 96 10 to 75 30 to 50 Protein 2 to 96  5 to 70 15 to 35Fat 2 to 96 20 to 85 35 to 55 Example J Example K Example L Carbohydrate25 to 50 25 to 50 35 to 50 Protein 10 to 30  5 to 30 7.5 to 25  Fat  1to 20  2 to 20 30 to 60

Protein

The infant formulas of the present disclosure may comprise protein inaddition to the micronutrients described herein. Any known or otherwisesuitable protein or protein source may be included in the infantformulas of the present disclosure, provided that such proteins aresuitable for feeding to infants, and in particular, newborn infants.

Non-limiting examples of suitable protein or sources thereof for use inthe infant formulas include hydrolyzed, partially hydrolyzed ornon-hydrolyzed proteins or protein sources, which may be derived fromany known or otherwise suitable source such as milk (e.g., casein,whey), animal (e.g., meat, fish), cereal (e.g., rice, corn), vegetable(e.g., soy), or combinations thereof. Non-limiting examples of suchproteins include milk protein isolates, milk protein concentrates asdescribed herein, casein protein isolates, extensively hydrolyzedcasein, whey protein, sodium or calcium caseinates, whole cow milk,partially or completely defatted milk, soy protein isolates, soy proteinconcentrates, and so forth. The proteins for use herein can alsoinclude, or be entirely or partially replaced by, free amino acids knownfor use in nutritional products, non-limiting examples of which includeL-alanine, L-aspartic acid, L-glutamic acid, glycine, L-histidine,L-isoleucine, L-leucine, L-phenylalanine, L-proline, L-serine,L-threonine, L-valine, L-tryptophan, L-glutamine, L-tyrosine,L-methionine, L-cysteine, taurine, L-arginine, L-carnitine, andcombinations thereof.

Fat

The infant formulas of the present disclosure may comprise a source orsources of fat in addition to micronutrients described herein. Suitablesources of fat for use in the infant formulas disclosed herein includeany fat or fat source that is suitable for use in an oral nutritionalproduct and is compatible with the essential elements and features ofsuch products, provided that such fats are suitable for feeding toinfants.

Non-limiting examples of suitable fats or sources thereof for use in theinfant formulas described herein include coconut oil, fractionatedcoconut oil, soybean oil, corn oil, olive oil, safflower oil, high oleicsafflower oil, high GLA-safflower oil, oleic acids, MCT oil (mediumchain triglycerides), sunflower oil, high oleic sunflower oil,structured triglycerides, palm and palm kernel oils, palm olein, canolaoil, flaxseed oil, borage oil, evening primrose oil, blackcurrant seedoil, transgenic oil sources, marine oils (e.g., tuna, sardine), fishoils, fungal oils, algae oils, cottonseed oils, and combinationsthereof. In one embodiment, suitable fats or sources thereof includeoils and oil blends including long chain polyunsaturated fatty acids(LC-PUFAs). Some non-limiting specific polyunsaturated acids forinclusion include, for example, docosahexaenoic acid (DHA), arachidonicacid (ARA), eicosapentaenoic acid (EPA), linoleic acid (LA), and thelike. Non-limiting sources of arachidonic acid and docosahexaenoic acidinclude marine oil, egg derived oils, fungal oil, algal oil, andcombinations thereof.

Carbohydrate

The infant formulas of the present disclosure may comprise anycarbohydrates that are suitable for use in an oral nutritional product,such as infant formula, and are compatible with the essential elementsand features of such product.

Non-limiting examples of suitable carbohydrates or sources thereof foruse in the infant formulas described herein may include maltodextrin,hydrolyzed, intact, or modified starch or cornstarch, glucose polymers,corn syrup, corn syrup solids, rice-derived carbohydrates, rice syrup,pea-derived carbohydrates, potato-derived carbohydrates, tapioca,sucrose, glucose, fructose, lactose, high fructose corn syrup, honey,sugar alcohols (e.g., maltitol, erythritol, sorbitol), artificialsweeteners (e.g., sucralose, acesulfame potassium, stevia), indigestibleoligosaccharides such as fructooligosaccharides (FOS), and combinationsthereof. In one embodiment, the carbohydrate may include a maltodextrinhaving a DE value of less than 20.

Other Optional Ingredients

The infant formulas of the present disclosure may further comprise otheroptional ingredients that may modify the physical, chemical, aestheticor processing characteristics of the products or serve as pharmaceuticalor additional nutritional components when used in the targetedpopulation. Many such optional ingredients are known or otherwisesuitable for use in medical food or other nutritional products orpharmaceutical dosage forms and may also be used in the compositionsherein, provided that such optional ingredients are safe for oraladministration and are compatible with the essential and otheringredients in the selected product form.

Non-limiting examples of such optional ingredients includepreservatives, anti-oxidants, emulsifying agents, buffers,fructooligosaccharides, galactooligosaccharides, human milkoligosaccharides and other prebiotics, pharmaceutical actives,additional nutrients as described herein, colorants, flavors, thickeningagents and stabilizers, emulsifying agents, lubricants, carotenoids(e.g., beta-carotene, zeaxanthin, lutein, lycopene), and so forth, andcombinations thereof.

A flowing agent or anti-caking agent may be included in the powderinfant formulas as described herein to retard clumping or caking of thepowder over time and to make a powder embodiment flow easily from itscontainer. Any known flowing or anti-caking agents that are known orotherwise suitable for use in a nutritional powder or product form aresuitable for use herein, non limiting examples of which includetricalcium phosphate, silicates, and combinations thereof. Theconcentration of the flowing agent or anti-caking agent in thenutritional product varies depending upon the product form, the otherselected ingredients, the desired flow properties, and so forth, butmost typically range from about 0.1% to about 4%, including from about0.5% to about 2%, by weight of the nutritional product.

A stabilizer may also be included in the infant formulas. Any stabilizerthat is known or otherwise suitable for use in a nutritional product isalso suitable for use herein, some non-limiting examples of whichinclude gums such as xanthan gum. The stabilizer may represent fromabout 0.1% to about 5.0%, including from about 0.5% to about 3%,including from about 0.7% to about 1.5%, by weight of the infantformula.

Stability

The low calorie, low micronutrient liquid infant formulas of the presentdisclosure advantageously exhibit improved physical attributes,including improved stability, as compared to low calorie, highmicronutrient formulas. Physical stability issues in liquid infantformulas often arise when the formulas are stored for extended periodsof time prior to use. During this time, components of the formulas, fatsfor example, often separate from the aqueous components. Components ofthe infant formula may also fall out of suspension, forming sediment atthe bottom of the formula container. Although this phase separation andsedimentation may be rectified by shaking the formula to remix formulacomponents, such phase separation and sedimentation often results ingreatly diminished consumer acceptance of the product.

It has now been discovered that the micronutrient content of low calorieliquid infant formulas may affect the stability of the infant formulas.In particular, the low calorie, low micronutrient liquid infant formulasof the present disclosure advantageously exhibit less sedimentation andless separation over the shelf life of the formulas, than do lowcalorie, high micronutrient formulas.

Protein Loading

A variety of measures may be used to demonstrate the stability of liquidinfant formulas. For instance, one way the stability of liquid infantformulas can be determined is by measuring the protein loading levels.Protein loading levels are expressed as the protein percent of a creamlayer formed following high speed centrifugation of the infant formula(the number of grams of protein per 100 grams of cream layer). Suitabletechniques for determining protein loading levels are described indetail in the examples of the current disclosure.

The stability of a liquid infant formula emulsion generally increaseswith increasing protein loading levels. It has now been discovered thatlow calorie, low micronutrient retort sterilized liquid infant formulashave higher levels of protein loading than low calorie, highmicronutrient retort sterilized liquid infant formulas. This was foundto be the case for both days 1-2 retort infant formulas and days 3-9retort infant formulas.

Thus, in one aspect, the present disclosure is directed to a lowcalorie, low micronutrient liquid infant formula having an increasedprotein loading level, as compared to a low calorie, high micronutrientinfant formula. Preferably, the low calorie, low micronutrient liquidinfant formula is a retort sterilized, ready-to-feed (RTF) formula. Inembodiments where the low calorie, low micronutrient liquid infantformula is a days 1-2 infant formula, the infant formula will typicallyhave a protein loading level of at least about 5.0%, including fromabout 5.0% to about 7.0%, from about 5.5% to about 6.5%, from about 5.7%to about 6.1%, and in particular about 5.9%.

In embodiments where the low calorie, low micronutrient liquid infantformula is a days 3-9 infant formula, the infant formula will typicallyhave a protein loading value of at least about 6.0%, including fromabout 6.0% to about 8.0%, from about 6.5% to about 7.5%, from about 6.7%to about 7.1%, and in particular about 6.9%. Preferably, the lowcalorie, low micronutrient liquid infant formula is retort sterilized.

Particle Size

Another measure that may be used to demonstrate the stability of liquidinfant formulas is particle size distribution and the average size ofparticles present in the infant formula. Particle size distribution andaverage particle size may be determined using any technique known in theart. One technique, described in the examples of the current disclosure,involves the use of a light scattering machine (e.g., Beckman Coulter LS13 320), which measures the size distribution of particles suspended ina sample of the liquid infant formula using multiple wavelength lightsources. Other suitable techniques may also be used.

Stability of a liquid infant formula emulsion generally increases withreducing particle size. It has now been discovered that the low calorie,low micronutrient days 1-2 retort sterilized liquid infant formulas ofthe present disclosure have a larger number of small particles, and asmaller average particle size for particles present in the formulas,than do low calorie, high micronutrient days 1-2 retort sterilizedliquid infant formulas.

Thus, in one aspect, the present disclosure is directed to a lowcalorie, low micronutrient liquid infant formula having a smalleraverage particle size for particles present in the formula, as comparedto a low calorie, high micronutrient liquid infant formula. Preferably,the low calorie, low micronutrient liquid infant formula is a retortsterilized RTF formula, and more preferably is a days 1-2 retortsterilized liquid infant formula. In embodiments where the low calorie,low micronutrient liquid infant formula is a days 1-2 infant formula,particles present in the infant formula will typically have an averageparticle size of from about 0.1 μm to about 1.0 μm, including from about0.15 μm to about 0.8 μm, and from about 0.15 μm to about 0.7 μm.

Typically, for the low calorie, low micronutrient days 1-2 liquid infantformulas of the present disclosure, at least about 50%, including fromabout 50% to about 100%, and from about 50% to about 70% of theparticles present in the infant formula will have a particle size(diameter) of from about 0.15 μm to about 0.8 μm.

Creaming Velocity

Another measure that may be used to demonstrate the stability of liquidinfant formulas is creaming velocity. Creaming velocity measures therate of movement of particles through a liquid sample, in this instance,an infant formula, and is predictive of the capacity of the infantformula to form a cream layer upon standing for extended periods of timeor upon centrifugation. Creaming velocity can be calculated using thefollowing equation:

$v_{cream} = {\frac{2}{9}\frac{\rho_{fluid} - \rho_{particle}}{\eta}{gR}^{2}}$

wherein:v_(cream) is the creaming velocityρ_(fluid) is the density of the formulaρ_(particle) is the density of the particlesη is the viscosity of the formulaR is the average particle sizeg is the gravitational acceleration

Stability of a liquid infant formula emulsion generally increases withdecreasing creaming velocity. It has now been discovered that the lowcalorie, low micronutrient days 1-2 retort sterilized liquid infantformulas of the present disclosure have a lower creaming velocity, thando low calorie, high micronutrient days 1-2 retort sterilized liquidinfant formulas.

Thus, in one aspect, the present disclosure is directed to a lowcalorie, low micronutrient liquid infant formula having a low creamingvelocity, as compared to a low calorie, high micronutrient infantformula. Preferably, the low calorie, low micronutrient liquid infantformula is a retort sterilized RTF formula, and more preferably is adays 1-2 retort sterilized liquid infant formula. In embodiments wherethe low calorie, low micronutrient liquid infant formula is a days 1-2infant formula, the infant formula will typically have a creamingvelocity about 5.0 cm/day or less, including from about 1.0 cm/day toabout 5.0 cm/day, from about 3.0 cm/day to about 3.5 cm/day, and inparticular about 3.2 cm/day.

Color

The low calorie, low micronutrient liquid infant formulas of the presentdisclosure also advantageously exhibit improved color, as compared tolow calorie, high micronutrient formulas.

Liquid infant formulas contain a variety of nutrients that potentiallyinteract during formulation, processing, and storage. Such interactionscan distort the color of the formula with gray, beige, or similar otherdiscolorations. Such discolorations often result in greatly diminishedacceptance of the product by consumers, who typically prefer a bright,whitish colored product.

One technique that can be used to evaluate the color characteristics ofan infant formula is Agtron color scores. Agtron scores as used hereinare measured by conventional techniques using an Agtron 45Spectrophotometer (available from Agtron Inc., Reno, Nev.). The Agtronscores are a measure of the percentage of reflected energy (light) fromthe surface of each infant formula. The more reflective or brighter incolor the formula surface, the higher the Agtron score. These scoresrange from zero (black) to 100 (white).

It has now been discovered that the micronutrient content of low calorieliquid infant formulas affects the color of the formulas. In particular,the low calorie, low micronutrient liquid infant formulas of the presentdisclosure advantageously have a brighter, whiter color, as defined byAgtron score, than do low calorie, high micronutrient formulas. This wasfound to be the case for both retort and aseptic low calorie, lowmicronutrient liquid formulas. The improved color of the low calorie,low micronutrient liquid infant formulas was also observed not just uponformulation, but also after extended periods of time, in some cases atleast 9 months following product formulation.

Thus, in one aspect, the present disclosure is directed to a lowcalorie, low micronutrient days 1-2 liquid infant formula that has anAgtron score following formulation (within a day of formulation) of atleast about 45, including from about 45 to about 60, and from about 47to about 55. Preferably, the formula is a retort sterilized RTF formula.In other embodiments, the formula has an Agtron score two months afterformulation of at least about 40, including from about 40 to about 50;has an Agtron score four months after formulation of at least about 37,including from about 40 to about 50; has an Agtron score six monthsafter formulation of at least about 37, including from about 37 to about50; and has an Agtron score nine months after formulation of at leastabout 35, including from about 35 to about 45.

In another aspect, the present disclosure is directed to a low calorie,low micronutrient days 3-9 liquid retort sterilized infant formula thathas an Agtron score following formulation of at least about 42,including from about 42 to about 55, and from about 45 to about 52. Inother embodiments, the formula has an Agtron score three months afterformulation of at least about 40, including from about 40 to about 50;and has an Agtron score six months after formulation of at least about40, including from about 40 to about 50.

In another aspect, the present disclosure is directed to a low calorie,low micronutrient days 3-9 liquid aseptic sterilized infant formula thathas an Agtron score following formulation of at least about 58,including from about 58 to about 65, and from about 60 to about 62. Inother embodiments, the formula has an Agtron score two months afterformulation of at least about 55, including from about 55 to about 62;has an Agtron score six months after formulation of at least about 55,including from about 55 to about 60; and has an Agtron score nine monthsafter formulation of at least about 52, including from about 52 to about55.

Buffering Capacity

The low calorie infant formulas of the present disclosure (having eithera high or a low micronutrient content) also advantageously exhibitimproved buffering capacity, as compared to full calorie formulas.

Human breast milk is believed to contain certain factors which promotethe development of a favorable intestinal bacterial flora, specifically,Bifidobacterium, which discourage the proliferation of pathogenicmicrobes. The growth of Bifidobacterium in the intestine of an infant isbelieved to be promoted by the physicochemical properties of humanbreast milk, particularly its high lactose content, which is a substratefor Bifidobacterium, its low protein content, and its low bufferingcapacity. Further, the low buffering capacity of human milk may allowthe natural level of acidity in gastrointestinal (GI) tract of infantsto be more effective in inactivating orally ingested pathogens. In somecases, infant formula may have a relatively high buffering capacity,which may not be completely favorable for the growth of Bifidobacterium,and may potentially impact the natural acidity of an infant's GI tract.Consequently, some formula fed infants may experience more episodes ofGI tract infection as compared to breast fed infants.

It has now been discovered that the buffering capacity of infant formulais correlated to the energy content of the formula. Specifically, it hasbeen discovered that the buffering capacity of infant formula decreaseswith decreasing energy content. The low calorie infant formulas of thepresent disclosure thus advantageously have an improved (i.e., lower)buffering capacity than full calorie infant formulas, and in someembodiments, have a lower buffering capacity than that of human milk.The low calorie infant formulas of the present disclosure can thus beused to regulate gastric acidity in infants, and in particular newborns,reduce the growth of pathogenic microorganisms in the infant GI tract,promote the growth of beneficial microorganisms, such asBifidobacterium, and increase the effectiveness of the inactivation oforally ingested pathogens.

Buffering capacity refers generally to the ability of a liquid to resistchanges in pH. There are several measures that can be used to expressbuffering capacity of the infant formulas of the present disclosure. Forinstance, buffering capacity of the infant formulas can be expressed asthe increase in hydrogen ion concentration ([H+]) following addition ofhydrochloric acid (HCl) to the infant formula (or to reconstitutedformula for powder infant formula embodiments). Specifically, bufferingcapacity can be expressed as the increase in [H+] following addition of5 mmoles of HCl to 100 mL of formula, or alternately, as the increase in[H+] following the addition of 5.50 mmoles of HCl to 100 mL of formula(or the addition of 2.75 mmoles of HCl to 50 mL of formula).

The low calorie infant formulas of the present disclosure may have abuffering capacity, expressed as the [H+] following addition of 5 mmolesof HCl to 100 mL of formula, of at least about 2.0 mM, including atleast about 5.0 mM, at least about 7.0 mM, at least about 10.0 mM, atleast about 13.0 mM, and at least about 17.0 mM, and/or from about 2.0mM to about 25.0 mM, including from about 5.0 mM to about 21.0 mM, andfrom about 10.0 mM to about 21.0 mM. The infant formulas may bereconstituted powder formulas, retort sterilized, or aseptic sterilized,and may be a days 1-2 or a days 3-9 formula. In one embodiment, the lowcalorie infant formula is a days 3-9 formula, and has a bufferingcapacity, expressed as the [H+] following addition of 5 mmoles of HCl to100 mL of formula at least about 2.0 mM, including at least about 5.0mM, at least about 7.0 mM, and at least about 9.0 mM, and/or from about2.0 mM to about 13.0 mM, including from about 8.0 mM to about 11.0 mM.In another embodiment, the low calorie infant formula is a days 1-2formula and has a buffering capacity, expressed as the [H+] followingaddition of 5 mmoles of HCl to 100 mL of formula, of at least about 8.0mM, including at least about 10.0 mM, at least about 13.0 mM, at leastabout 17.0 mM, and at least about 20.0 mM, and/or from about 8.0 mM toabout 25.0 mM, including from about 8.0 mM to about 21.0 mM, from about13.0 mM to about 20.0 mM, and from about 17.0 mM to about 20.0 mM.

Alternately, the buffering capacity of the infant formula can beexpressed as the decrease in pH of the formula following addition of HClto the infant formula (or to reconstituted formula for powder infantformula embodiments). Specifically, buffering capacity can be expressedas the decrease in pH following addition of 5.50 mmoles of HCl to 100 mLof formula (or the addition of 2.75 mmoles of HCl to 50 mL of formula).

Thus, in one embodiment, the low calorie infant formulas of the presentdisclosure is a powder infant formula, and may have a buffering capacityfollowing reconstitution, expressed as the decrease in pH of the formulafollowing addition of 5.50 mmoles of HCl to 100 mL of reconstitutedformula, of at least about 4.20, including at least about 4.50, and atleast about 4.80. In another embodiment where the low calorie infantformula is a retort sterilized RTF formula, the buffering capacity,expressed as the decrease in pH of the formula following addition of2.75 mmoles of HCl to 50 mL of formula, is at least about 4.20,including at least about 4.30. In still another embodiment wherein thelow calorie infant formula is an aseptic sterilized RTF formula, thebuffering capacity, expressed as the decrease in pH of the formulafollowing addition of 5.50 mmoles of HCl to 100 mL of formula, is atleast about 4.60, including at least about 4.70.

Another measure of buffering capacity is buffering strength. Unlessotherwise indicated, the buffering strength of the infant formulas ofthe present disclosure is expressed as the volume of 0.1 M HCl needed todecrease the pH of 50 mL of formula (or reconstituted formula for powderinfant formula embodiments) from the starting pH (e.g., 6.0) to a pH of3.0. As used herein, the term “low buffering strength” refers to abuffering strength of about 18 mL or less. Buffering strength is alsoexpressed herein (where indicated) as mmoles of HCl required to lowerthe pH of 100 mL of formula from 6.0 to 3.0 and as mmoles of HClrequired to lower the pH of 50 mL of formula from 6.0 to 3.0.

The low calorie infant formulas of the present disclosure may have abuffering strength, expressed as the mL of 0.1 M HCl needed to decreasethe pH of 50 mL of formula (or reconstituted formula for powder infantformula embodiments) from the starting pH to a pH of 3.0, of about 18 mLor less, including about 14 mL or less, and/or including from about 9 mLto about 18 mL, including from about 10 mL to about 14 mL, and fromabout 14 mL to about 18 mL. In one embodiment, the low calorie infantformula is a days 3-9 formula, and has a buffering strength of about 18mL or less, including from about 14 mL to about 18 mL, and from about 16mL to about 17 mL. In another embodiment, the low calorie infant formulais a days 1-2 formula, and has a buffering strength of about 14 mL orless, including from about 9 mL to about 14 mL, and from about 10 mL toabout 11 mL. The buffering strength of human milk typically ranges from9 mL to 18 mL. The low calorie infant formulas of the present disclosureadvantageously have a buffering strength comparable to or lower thanthat of human milk.

Protein Hydrolysis and Digestion

The low calorie infant formulas of the present disclosure (having eithera high or a low micronutrient content) also advantageously exhibit afaster rate of protein hydrolysis and digestion, as compared to fullcalorie formulas.

Two factors in determining the nutritional quality of food proteins aredigestibility and bioavailability. Typically, infant formulas contain ahigher level of protein than the level found in breast milk. Infantformulas are typically manufactured with higher levels of proteins toaccount for the assumed lower digestibility of the proteins.

Further, in some cases, the processes used during the manufacture ofinfant formulas may potentially have some nutritional consequences, suchas lowered solubility and/or digestibility of the proteins in theformula. For example, some heat treatments over extended periods of timethat are used to produce concentrated liquid and ready-to-feed infantformulas may possibly decrease digestibility of proteins in some cases.As a result of exposure to heat, proteins denature or aggregate,possibly altering their digestibility in some cases. The treatment ofmilk at high temperatures may also increase reactions of amino acidswith sugars known as Maillard reactions. These reactions may decreasethe bioavailability of amino acids by limiting the accessibility ofproteolytic enzymes in some cases. As a result, some formula fed infantsmay potentially experience some incomplete nutrient (and in particularprotein) absorption. Consequently, an infant formula having improvedprotein digestion would be beneficial, especially for newborn infantswho are known to have lower amounts of digestive enzymes, such a gastricpepsin and intestinal pancreatin, than do older infants and adults.

It has now been discovered that the extent (used interchangeably hereinwith the term “rate”) of digestion (used interchangeably herein with theterm “hydrolysis”) of protein in infant formula is correlated to theenergy content of the formula. Specifically, it has been discovered thatthe rate of digestion of protein present in the infant formula increaseswith decreasing energy content of the formula. The low calorie infantformulas of the present disclosure thus advantageously have an improved(e.g., faster) rate of protein digestion than do full calorie infantformulas. This may result in improved infant formula tolerance andimproved nutrient (and in particular protein) absorption by the infant.

There are several measures that can be used to express the rate orextent of protein digestion. For instance, the rate or extent ofdigestion of the proteins in the infant formulas of the presentdisclosure can be expressed as the median molecular weight (MW) of theproteins following an in vitro gastrointestinal digestion using pepsinand pancreatin (amylase/protease/lipase) or an in vitro pancreatindigestion. A decreasing protein MW median is indicative of a faster rateand increased extent of digestion. The procedures for these digestionsare set forth in the examples.

In some embodiments, the low calorie infant formulas of the presentdisclosure may have a rate or extent of protein digestion, expressed asthe protein MW median following in vitro gastrointestinal digestion,performed as described herein, of about 950 Daltons (Da) or less,including about 925 Da or less, about 850 Da or less, about 800 Da orless, and about 790 Da or less. For days 3-9 formulas of the presentdisclosure, the rate or extent of protein digestion, expressed as theprotein MW median following in vitro gastrointestinal digestion,performed as described herein, is typically from about 700 Da to about950 Da. For days 1-2 formulas, the rate or extent of protein digestion,expressed as the protein MW median following in vitro gastrointestinaldigestion, performed as described herein, is typically about 825 Da orless, including about 800 Da or less, about 780 Da or less, about 750 Daor less and about 720 Da or less. Typically the rate or extent ofprotein digestion for days 1-2 formulas is from about 700 Da to about800 Da.

The low calorie infant formulas of the present disclosure may have arate or extent of protein digestion, expressed as the protein MW medianfollowing in vitro pancreatin digestion for 71 minutes, performed asdescribed herein, of about 800 Da or less, including about 775 Da orless, and about 750 Da or less, and in particular from about 725 Da toabout 775 Da for days 3-9 formulas. For days 1-2 formulas, the rate orextent of protein digestion, expressed as the protein MW medianfollowing in vitro pancreatin digestion for 71 minutes, performed asdescribed herein, is typically about 750 Da or less, including about 725Da or less, about 700 Da or less, and about 690 Da or less, and inparticular from about 675 Da or less to about 700 Da or less.

The low calorie infant formulas of the present disclosure may have arate or extent of protein digestion, expressed as the protein MW medianfollowing in vitro pancreatin digestion for 60 minutes, performed asdescribed herein, of about 1000 Da or less, including about 950 Da orless, about 900 Da or less, about 850 Da or less, about 825 Da or less,and about 810 Da or less, and in particular from about 775 Da to about825 Da.

The rate or extent of protein digestion can also be expressed as thepercent of total proteins having a MW of greater than 5000 Da, followingeither the in vitro gastrointestinal digestion or the in vitropancreatin digestion described herein. A smaller percentage isindicative of a faster rate and increased extent of digestion. The lowcalorie infant formulas of the present disclosure may have a rate orextent of protein digestion, expressed as the percent of total proteinshaving a MW of greater than 5000 Da following in vitro gastrointestinaldigestion, performed as described herein, of about 13.5% or less,including about 12.0% or less, about 11.0% or less, about 9.0% or less,and about 6.0% or less, and in particular from about 5.0% to about 13.5%for powder formulas. In embodiments where the infant formula is retortsterilized, the rate or extent of protein digestion, expressed as thepercent of total proteins having a MW of greater than 5000 Da followingin vitro gastrointestinal digestion, performed as described herein, isabout 8.0% or less, including about 7.0% or less, about 6.0% or less,about 5.0% or less, about 4.0% or less, and about 3.0% or less, andfurther including from about 2.0% to about 6.0%. In embodiments wherethe infant formula is aseptic sterilized, the rate or extent of proteindigestion, expressed as the percent of total proteins having a MW ofgreater than 5000 Da following in vitro gastrointestinal digestion,performed as described herein, is about 9.0% or less, including about7.0% or less, about 6.0% or less, about 5.0% or less, about 3.0% orless, and further including from about 2.0% to about 5.0%.

The rate or extent of protein digestion can also be expressed by theamount of insoluble protein present in the infant formula following invitro gastrointestinal digestion, performed as described herein.Techniques for determining the level of insoluble protein are set forthin the examples of the present disclosure. A smaller amount of insolubleprotein is indicative of a faster rate and increased extent ofdigestion.

The low calorie infant formulas of the present disclosure may have arate or extent of protein digestion, expressed as the amount ofinsoluble protein in the formula following in vitro gastrointestinaldigestion, performed as described herein, of about 150 mg/L or less,including about 110 mg/L or less, about 75 mg/L or less, about 50 mg/Lor less, and about 25 mg/L or less, and in particular from about 20 mg/Lto about 110 mg/L.

As discussed herein, processing of infant formulas, and in particularthe treatment of milk products at high temperatures may increasereactions of amino acids with sugars, known as Maillard reactions. Thesereactions decrease the bioavailability of amino acids by limiting theaccessibility of proteolytic enzymes. It has now been discovered thatMaillard reactions proceed to a lesser extent in the low calorie infantformulas of the present disclosure as compared to full calorie formulas.This may be illustrated by determining the level of Maillard reactionmarkers in the infant formula following digestion. Specifically, the lowcalorie infant formulas of the present disclosure have been found tohave lower levels of the Maillard reaction marker furosine, following invitro gastrointestinal digestion performed as described herein, than dofull calorie formulas.

Thus, in one aspect the present disclosure provides infant formulas thatcomprise, following in vitro gastrointestinal digestion performed asdescribed herein, the Maillard reaction marker furosine in amounts(mg/100 g product) of about 2.5 or less, including about 1.5 or less,about 1.0 or less, and about 0.90 or less, and in particular from about0.7 to about 1.0.

Methods of Manufacture

The infant formulas of the present disclosure may be prepared by anyknown or otherwise effective manufacturing technique for preparing theselected product solid or liquid form. Many such techniques are knownfor any given product form such as nutritional liquids or powders andcan easily be applied by one of ordinary skill in the art to the infantformulas described herein.

The infant formulas of the present disclosure can therefore be preparedby any of a variety of known or otherwise effective formulation ormanufacturing methods. In one suitable manufacturing process, forexample, at least two separate slurries are prepared, that are laterblended together, heat treated, standardized, and either terminallysterilized to form a retort infant formula or aseptically processed andfilled to form an aseptic infant formula. Alternately, the slurries canbe blended together, heat treated, standardized, heat treated a secondtime, evaporated to remove water, and spray dried to form a powderinfant formula.

The slurries formed may include a carbohydrate-mineral (CHO-MIN) slurryand a protein-in-oil (PIO) slurry. Initially, the CHO-MN slurry isformed by dissolving selected carbohydrates (e.g., lactose,galactooligosaccharides, etc.) in heated water with agitation, followedby the addition of minerals (e.g., potassium citrate, magnesiumchloride, potassium chloride, sodium chloride, choline chloride, etc.).The resulting CHO-MIN slurry is held with continued heat and moderateagitation until it is later blended with the other prepared slurries.

The PIO slurry is formed by heating and mixing the oil (e.g., high oleicsafflower oil, soybean oil, coconut oil, monoglycerides, etc.) andemulsifier (e.g., soy lecithin), and then adding oil soluble vitamins,mixed carotenoids, protein (e.g., milk protein concentrate, milk proteinhydrolysate, etc.), carrageenan (if any), calcium carbonate ortricalcium phosphate (if any), and ARA oil and DHA oil (in someembodiments) with continued heat and agitation. The resulting PIO slurryis held with continued heat and moderate agitation until it is laterblended with the other prepared slurries.

Water was heated and then combined with the CHO-MIN slurry, nonfat milk(if any), and the PIO slurry under adequate agitation. The pH of theresulting blend was adjusted to 6.6-7.0, and the blend was held undermoderate heated agitation. ARA oil and DHA oil is added at this stage insome embodiments.

The composition is then subjected to high-temperature short-time (HTST)processing, during which the composition is heat treated, emulsified andhomogenized, and then cooled. Water soluble vitamins and ascorbic acidare added, the pH is adjusted to the desired range if necessary, flavors(if any) are added, and water is added to achieve the desired totalsolid level. For aseptic infant formulas, the emulsion receives a secondheat treatment through an aseptic processor, is cooled, and thenaseptically packaged into suitable containers. For retort infantformulas, the emulsion is packaged into suitable containers andterminally sterilized. In some embodiments, the emulsions can beoptionally further diluted, heat-treated, and packaged to form a desiredready-to-feed or concentrated liquid, or can be heat-treated andsubsequently processed and packaged as a reconstitutable powder, e.g.,spray dried, dry mixed, agglomerated.

The spray dried powder infant formula or dry-mixed powder infant formulamay be prepared by any collection of known or otherwise effectivetechniques, suitable for making and formulating a nutritional powder.For example, when the powder infant formula is a spray-dried nutritionalpowder, the spray drying step may likewise include any spray dryingtechnique that is known for or otherwise suitable for use in theproduction of nutritional powders. Many different spray drying methodsand techniques are known for use in the nutrition field, all of whichare suitable for use in the manufacture of the spray dried powder infantformulas herein. Following drying, the finished powder may be packagedinto suitable containers.

Methods of Use

The low calorie infant formulas of the present disclosure may be orallyadministered to infants, including term, preterm, and/or newborninfants. The low calorie infant formulas may be administered as a sourceof nutrition for infants and/or can be used to address one or more ofthe diseases or conditions discussed herein, or can be used to provideone or more of the benefits described herein, to preterm infants, terminfants, and/or newborn infants. Any of this group may actually have thedisease or condition, or may be at risk of getting the disease orcondition (due to family history etc.), may be susceptible to thedisease or condition, or may be in need of treatment/control/reductionof a certain disease or condition. The infant formulas will typically beadministered daily, at intake volumes suitable for the age of theinfant. As such, because some of the method embodiments disclosed hereinare directed to certain subsets or subclasses of infants (e.g., thoseinfants in need of treatment or control of a disease or condition) andnot generally to the standard infant population, not all infants canbenefit from all method embodiments disclosed herein.

For instance, the methods of the present disclosure may includeadministering one or more of the low calorie formulas of the presentdisclosure to an infant at the average intake volumes described herein.In some embodiments, newborn infants are provided with increasingformula volumes during the initial weeks of life. Such volumes mosttypically range up to about 100 mL/day on average during the first dayor so of life; up to about 200 to about 700 mL/day, including from about200 to about 600 mL/day, and also including from about 250 to about 500mL/day, on average during the remainder of the three month newbornfeeding period. It is to be understood, however, that such volumes canvary considerably depending upon the particular newborn infant and theirunique nutritional needs during the initial weeks or months of life, aswell as the specific nutrients and caloric density of the infant formulaadministered.

In some embodiments, the methods of the present disclosure may bedirected to newborn infants during the initial weeks or months of life,preferably during at least the first week of life, more preferablyduring at least the first two weeks of life, and including up to about 3months of life. Thereafter, the infant may be switched to a conventionalinfant formula, alone or in combination with human milk.

The methods described herein may comprise administering two or moredifferent infant formulas to the infant. For instance, the infant may beadministered a low calorie days 1-2 infant formula during the first twodays following birth and may then subsequently be administered a lowcalorie days 3-9 infant formula on days 3 to 9 following birth.Optionally, the days 3-9 infant formula may be administered past day 9following birth, or alternatively, a higher calorie formula (Includingfull calorie formulas) may be administered starting on day 10 followingbirth.

The infant formulas used in the methods described herein, unlessotherwise specified, are nutritional formulas and may be in any productform, including ready-to-feed liquids, concentrated liquids,reconstituted powders, and the like. In embodiments where the infantformulas are in powder form, the method may further comprisereconstituting the powder with an aqueous vehicle, most typically wateror human milk, to form the desired caloric density, which is then orallyor enterally fed to the infant. The powdered formulas are reconstitutedwith a sufficient quantity of water or other suitable fluid such ashuman milk to produce the desired caloric density, as well as thedesired feeding volume suitable for one infant feeding. The infantformulas may also be sterilized prior to use through retort or asepticmeans.

Other embodiments are described in more detail below.

Nutrition

In one aspect, the present disclosure is directed to a method ofproviding nutrition to an infant. The method comprises administering tothe infant any one or more of the low calorie, low micronutrient infantformulas of the present disclosure. Such methods may include the dailyadministration of the infant formulas, including administration at thedaily intake volumes as described hereinbefore. In some embodiments, theinfant is a newborn infant.

As noted above, any of the low calorie, low micronutrient infantformulas of the present disclosure may be used in this method.Specifically, the low micronutrient infant formula comprisesmicronutrients and at least one macronutrient selected from the groupconsisting of protein, carbohydrate, fat, and combinations thereof. Inone embodiment, the low micronutrient infant formula has an energycontent of from about 200 kcal/L to less than 600 kcal/L, wherein atleast 65% of the micronutrients are included in the infant formula in anamount that is from about 30% to about 80% of conventional amounts ofcorresponding micronutrients, on a per volume basis. In anotherembodiment, the low micronutrient infant formula has an energy contentof from about 200 kcal/L to about 360 kcal/L, wherein at least 45% ofthe micronutrients are included in the infant formula in an amount thatis from about 30% to about 65% of conventional amounts of correspondingmicronutrients, on a per volume basis. In still another embodiment, thelow micronutrient infant formula has an energy content of from about 360kcal/L to less than 600 kcal/L, wherein at least 30% of themicronutrients are included in the infant formula in an amount that isfrom about 55% to about 80% of conventional amounts of correspondingmicronutrients, on a per volume basis. The low calorie infant formulamay be a days 1-2 and/or a days 3-9 formula.

The method may also further comprise administering two or more differentinfant formulas to the infant. For instance, in one embodiment, theinfant is administered a low calorie infant formula (having either ahigh or low micronutrient content) having an energy content of fromabout 200 kcal/L to about 360 kcal/L (e.g., a days 1-2 formula) duringthe first two days following birth, and is subsequently administered alow calorie infant formula (having either a high or low micronutrientcontent) having an energy content of from about 360 kcal/L to less than600 kcal/L (e.g., a days 3-9 formula) on days 3 to 9 following birth.Optionally, the days 3-9 formula may be administered past day 9following birth, or alternatively, a higher calorie formula (Includingfull calorie formulas) may be administered starting on day 10 followingbirth.

Buffering Capacity

It has been discovered that the buffering capacity of infant formula iscorrelated to the energy content of the formula. Specifically, it hasbeen discovered that the buffering capacity of infant formula decreaseswith decreasing energy content. The low calorie infant formulas of thepresent disclosure thus advantageously have an improved (i.e., lower)buffering capacity than full calorie infant formulas, and in someembodiments, have a lower buffering capacity than human breast milk. Thelow calorie infant formulas of the present disclosure can thus be usedto increase the level of gastric acidity in infants, and in particularnewborns, and to regulate the growth of gastrointestinal flora ininfants, including controlling (e.g., reducing) the growth of pathogenicmicroorganisms in the infant GI tract, promoting the growth ofbeneficial microorganisms in the infant GI tract, and increasing theeffectiveness of the inactivation of orally ingested pathogens.

Without wishing to be bound to any particular theory, it is believedthat the more acidic pH in the GI tract of breastfed infants, ascompared to infants fed full calorie formulas, helps inactivate orallyingested pathogens, and provides a more hospitable environment for thegrowth of naturally occurring beneficial gastrointestinal flora. This isbelieved to be due, at least in part, to the low buffering capacity ofhuman breast milk. Because the low calorie infant formulas of thepresent disclosure have a buffering capacity comparable to or lower thanthat of human breast milk, infants fed the low calorie infant formulasdisclosed herein will have a level of gastric acidity more closelyresembling that found in breastfed infants.

Thus, in one aspect, the present disclosure is directed to a method forincreasing the level of gastric acidity (e.g., by lowering gastric pH)in an infant to about the same level of a breastfed infant. The methodcomprises identifying an infant having a depressed level of gastricacidity, and administering to the infant any of the low calorie infantformulas of the present disclosure. Preferably, the infant is a newborninfant.

The term “level of gastric acidity” refers to the level of acidity inthe stomach, and can be measured using pH. For instance, as the pH ofthe gastric contents decreases, the level of gastric acidity increases.As used herein, the term “depressed level of gastric acidity” means thelevel of gastric acidity in the infant is lower than that typicallyfound in breastfeed infants. Infants having a depressed level of gastricacidity can be identified as having a reduced or lower rate ofpathogenic bacteria colonization in the gut. Upon administration of thelow calorie infant formula of the present disclosure, the level ofgastric acidity in the infant is increased to the levels typically foundin breastfed infants.

As noted above, any of the low calorie infant formulas of the presentdisclosure may be used in this method. The low calorie infant formulamay have a low micronutrient content, or, in some embodiments, may havea high micronutrient content, and may be a days 1-2 or a days 3-9formula. In one embodiment, the infant formula has an energy content offrom about 200 kcal/L to about 500 kcal/L.

The method may also further comprise administering two or more differentinfant formulas to the infant. For instance, in one embodiment, theinfant is administered a days 1-2 formula having an energy content offrom about 200 kcal/L to about 360 kcal/L during the first two daysfollowing birth, and is subsequently administered a days 3-9 formulahaving an energy content of from about 360 kcal/L to less than 600kcal/L on days 3 to 9 following birth. Optionally, the days 3-9 formulamay be administered past day 9 following birth, or alternatively, ahigher calorie formula (Including full calorie formulas) may beadministered starting on day 10 following birth. The formula (s)administered to the infant will typically be administered daily atintake volumes as described hereinbefore.

In another aspect, the present disclosure is directed to a method forincreasing the level of gastric acidity in an infant comprisingadministering to the infant any of the low micronutrient infant formulasof the present disclosure. Preferably, the infant is a newborn infant.The low micronutrient infant formula comprises micronutrients and atleast one macronutrient selected from the group consisting of protein,carbohydrate, fat, and combinations thereof. In one embodiment, the lowmicronutrient infant formula has an energy content of from about 200kcal/L to less than 600 kcal/L, wherein at least 65% of themicronutrients are included in the infant formula in an amount that isfrom about 30% to about 80% of conventional amounts of correspondingmicronutrients, on a per volume basis. In another embodiment, the lowmicronutrient infant formula has an energy content of from about 200kcal/L to about 360 kcal/L, wherein at least 45% of the micronutrientsare included in the infant formula in an amount that is from about 30%to about 65% of conventional amounts of corresponding micronutrients, ona per volume basis. In still another embodiment, the low micronutrientinfant formula has an energy content of from about 360 kcal/L to lessthan 600 kcal/L, wherein at least 30% of the micronutrients are includedin the infant formula in an amount that is from about 55% to about 80%of conventional amounts of corresponding micronutrients, on a per volumebasis. The low calorie infant formula may be a days 1-2 and/or a days3-9 formula.

These methods may also further comprise administering two or moredifferent infant formulas to the infant. For instance, in oneembodiment, the infant is administered a low calorie infant formula(having either a high or low micronutrient content) having an energycontent of from about 200 kcal/L to about 360 kcal/L (e.g., a days 1-2formula), during the first two days following birth. The infant may thensubsequently be administered a low calorie infant formula (having eithera high or low micronutrient content) that has an energy content of fromabout 360 kcal/L to less than 600 kcal/L (e.g., a days 3-9 formula) ondays 3 to 9 following birth. Optionally, the days 3-9 formula may beadministered past day 9 following birth, or alternatively, a highercalorie formula (Including full calorie formulas) may be administeredstarting on day 10 following birth. In embodiments where the low calorieinfant formulas have a low micronutrient content, the amounts ofmicronutrients included in the formulas may be any of those set forthabove. The formula (s) administered to the infant will typically beadministered daily at intake volumes as described hereinbefore.

In still another embodiment, the present disclosure is directed to amethod for regulating growth of beneficial gastrointestinal flora in aninfant. The method comprises identifying an infant having an imbalancein the growth of gastrointestinal flora, and administering to the infantany of the low calorie infant formulas of the present disclosure.Preferably, the infant is a newborn infant.

For purposes of the present disclosure, the growth of gastrointestinalflora can be regulated by either promoting the growth of microorganismsbeneficial to GI health, and/or by controlling the growth of pathogenicmicroorganisms. The growth of pathogenic microorganisms can becontrolled by suppressing, inhibiting, killing, inactivating, destroyingor otherwise interfering with the growth of the pathogenicmicroorganisms, such that the growth rate of these microorganisms isslowed or stopped. Infants having an imbalance in the growth of GI florainclude infants in which the levels of one or more pathogenicmicroorganism in the infant's GI tract is higher than the levelstypically found in breastfed infants and/or the levels of one or morebeneficial microorganism in the infant's GI tract are lower than thelevels typically found in breastfeed infants. Such infants may beidentified by a lower rate of pathogenic bacteria colonization in thegut. Upon administration of the low calorie infant formula of thepresent disclosure, the level of gastric acidity in the infant isincreased to the levels similar to those typically found in breastfedinfants, resulting in a GI environment which promotes the growth ofbeneficial microorganisms and controls the growth of pathogenicmicroorganisms.

As noted above, any of the low calorie infant formulas of the presentdisclosure may be used in this method. The low calorie infant formulamay have a low micronutrient content, or, in some embodiments, may havea high micronutrient content, and may be a days 1-2 or a days 3-9formula. In one embodiment, the infant formula has an energy content offrom about 200 kcal/L to about 500 kcal/L of formula.

The method may also further comprise administering two or more differentinfant formulas to the infant. For instance, in one embodiment, theinfant is administered a days 1-2 formula having an energy content offrom about 200 kcal/L to about 360 kcal/L during the first two daysfollowing birth, and is subsequently administered a days 3-9 formulahaving an energy content of from about 360 kcal/L to less than 600kcal/L on days 3 to 9 following birth. Optionally, the days 3-9 formulamay be administered past day 9 following birth, or alternatively, ahigher calorie formula (Including full calorie formulas) may beadministered starting on day 10 following birth. The formula (s)administered to the infant will typically be administered daily atintake volumes as described hereinbefore.

In another aspect, the present disclosure is directed to a method forregulating the growth of gastrointestinal flora in an infant comprisingadministering to the infant any of the low micronutrient infant formulasof the present disclosure. Preferably, the infant is a newborn infant.The low micronutrient infant formula may be any of those set forthabove.

These methods may also further comprise administering two or moredifferent infant formulas to the infant. For instance, in oneembodiment, the infant is administered a low calorie infant formula(having either a high or low micronutrient content) having an energycontent of from about 200 kcal/L to about 360 kcal/L (e.g., a days 1-2formula), during the first two days following birth. The infant may thensubsequently be administered a low calorie infant formula (having eithera high or low micronutrient content) that has an energy content of fromabout 360 kcal/L to less than 600 kcal/L (e.g., a days 3-9 formula) ondays 3 to 9 following birth. Optionally, the days 3-9 formula may beadministered past day 9 following birth, or alternatively, a highercalorie formula (Including full calorie formulas) may be administeredstarting on day 10 following birth. In embodiments where the low calorieinfant formulas have a low micronutrient content, the amounts ofmicronutrients included in the formulas may be any of those set forthabove. The formula (s) administered to the infant will typically beadministered daily at intake volumes as described hereinbefore.

Beneficial microorganisms refer to those microorganisms that maintainthe microbial ecology of the GI tract, and show physiological,immuno-modulatory, and/or antimicrobial effects, such that theirpresence has been found to prevent and treat GI diseases and/ordisorders. Non-limiting examples of beneficial microorganisms includeany one or more of the following: the genus Lactobacillus including L.acidophilus, L. amylovorus, L. brevis, L. bulgaricus, L. casei spp.Casei, L. casei spp. Rhamnosus, L. crispatus, L. delbrueckii ssp.Lactis, L. fermentum, L. helvaticus, L. johnsonii, L. paracasei, L.pentosus, L. plantarum, L. reuteri, and L. sake; the genusBifidobacterium including B. animalis, B. bifidum, B. breve, B.infantis, and B. longum; the genus Pediococcus including P.acidilactici; the genus Propionibacterium including P. acidipropionici,P. freudenreichii, P. jensenii, and P. theonii; and the genusStreptococcus including S. cremoris, S. lactis, and S. thermophilus; andcombinations thereof.

Non-limiting examples of pathogenic microorganisms whose growth may becontrolled by the methods disclosed herein include any one or more ofthe following: bacteria such as the genus Clostridum including C.difficile; Escherichia coli (E. coli); Vibrio sp.; Salmonella sp.;Shigella sp.; Camphylobacter sp.; Aeromonas sp.; Staphylococcus sp.;Pseudomonas sp.; and parasites such as Giardia sp.; and Cryptosporidiumsp.; and combinations thereof.

Protein Digestion and Hydrolysis

It has been discovered that the rate and extent of digestion of proteinin infant formula is correlated to the energy content of the formula.Specifically, it has been discovered that the rate of digestion ofproteins in infant formula increases with decreasing energy content ofthe formula. The low calorie infant formulas of the present disclosurethus advantageously have an improved (e.g., faster) rate of digestion ascompared to full calorie infant formulas. The low calorie infantformulas of the present disclosure can thus be used to improve formulatolerance, protein digestion, and nutrient (and in particular protein)absorption in infants, and in particular newborns.

Thus, in one aspect, the present disclosure is directed to a method forimproving protein digestion in an infant. The method comprisesidentifying an infant experiencing incomplete protein digestion, andadministering to the infant any of the low calorie infant formulas ofthe present disclosure. Preferably, the infant is a newborn infant.

As used herein, the term “improving protein digestion” includesincreasing the rate of digestion (or hydrolysis) of protein present inthe infant formula and/or increasing the extent to which protein in theinfant formula is digested when contacted with digestive enzymes. Thisimprovement in protein digestion can be determined using any of themeasures described herein, including, for example, the protein medianweight following digestion, the percent of total protein having amolecular weight of greater than 5000 Daltons following digestion,and/or the amount of insoluble protein present in the formula followingdigestion.

As used herein, the term “incomplete protein digestion” means the amountof protein, present in nutritional products consumed by the infant, thatis actually digested is lower than the amount typically digested bybreastfed infants. Infants experiencing incomplete protein digestion mayshow signs of formula intolerance, and may thus be identified using anyof the symptoms of formula intolerance described herein. Infantsexperiencing incomplete protein digestion can also be identified bydiarrhea, loose stools, gas, and/or bloating. Upon administration of alow calorie infant formula of the present disclosure, the rate andextent of protein digestion is improved.

As noted above, any of the low calorie infant formulas of the presentdisclosure may be used in this method. The low calorie infant formulamay have a low micronutrient content, or, in some embodiments, may havea high micronutrient content, and may be a days 1-2 and/or a days 3-9formula. In one embodiment, the infant formula has an energy content offrom about 200 kcal/L to less than 600 kcal/L of formula.

The method may also further comprise administering two or more differentinfant formulas to the infant. For instance, in one embodiment, theinfant is administered a days 1-2 formula having an energy content offrom about 200 kcal/L to about 360 kcal/L during the first two daysfollowing birth, and is subsequently administered a days 3-9 formulahaving an energy content of from about 360 kcal/L to less than 600kcal/L on days 3 to 9 following birth. Optionally, the days 3-9 formulamay be administered past day 9 following birth, or alternatively, ahigher calorie formula (Including full calorie formulas) may beadministered starting on day 10 following birth. The formula (s)administered to the infant will typically be administered daily atintake volumes as described hereinbefore.

In another aspect, the present disclosure is directed to a method forimproving protein digestion in an infant comprising administering to theinfant any of the low micronutrient infant formulas of the presentdisclosure. Preferably, the infant is a newborn infant. The lowmicronutrient infant formula comprises micronutrients and at least onemacronutrient selected from the group consisting of protein,carbohydrate, fat, and combinations thereof. In one embodiment, the lowmicronutrient infant formula has an energy content of from about 200kcal/L to less than 600 kcal/L, wherein at least 65% of themicronutrients are included in the infant formula in an amount that isfrom about 30% to about 80% of conventional amounts of correspondingmicronutrients, on a per volume basis. In another embodiment, the lowmicronutrient infant formula has an energy content of from about 200kcal/L to about 360 kcal/L, wherein at least 45% of the micronutrientsare included in the infant formula in an amount that is from about 30%to about 65% of conventional amounts of corresponding micronutrients, ona per volume basis. In still another embodiment, the low micronutrientinfant formula has an energy content of from about 360 kcal/L to lessthan 600 kcal/L, wherein at least 30% of the micronutrients are includedin the infant formula in an amount that is from about 55% to about 80%of conventional amounts of corresponding micronutrients, on a per volumebasis. The low calorie infant formula may be a days 1-2 and/or a days3-9 formula.

These methods may also further comprise administering two or moredifferent infant formulas to the infant. For instance, in oneembodiment, the infant is administered a low calorie infant formula(having either a high or low micronutrient content) having an energycontent of from about 200 kcal/L to about 360 kcal/L (e.g., a days 1-2formula), during the first two days following birth. The infant may thensubsequently be administered a low calorie infant formula (having eithera high or low micronutrient content) that has an energy content of fromabout 360 kcal/L to less than 600 kcal/L (e.g., a days 3-9 formula) ondays 3 to 9 following birth. Optionally, the days 3-9 formula may beadministered past day 9 following birth, or alternatively, a highercalorie formula (Including full calorie formulas) may be administeredstarting on day 10 following birth. In embodiments where the low calorieinfant formulas have a low micronutrient content, the amounts ofmicronutrients included in the formulas may be any of those set forthabove. The formula (s) administered to the infant will typically beadministered daily at intake volumes as described hereinbefore.

In still another embodiment, the present disclosure is directed to amethod of improving protein absorption in an infant. The methodcomprises identifying an infant experiencing incomplete proteinabsorption; and administering to the infant any of the low calorieinfant formulas of the present disclosure. Infants experiencingincomplete protein absorption may be identified using any of thecriteria described herein for identifying infants experiencingincomplete protein digestion.

As noted above, any of the low calorie infant formulas of the presentdisclosure may be used in this method. The low calorie infant formulamay have a low micronutrient content, or, in some embodiments, may havea high micronutrient content, and may be a days 1-2 or a days 3-9formula. In one embodiment, the infant formula has an energy content offrom about 200 kcal/L to less than 600 kcal/L of formula.

The method may also further comprise administering two or more differentinfant formulas to the infant. For instance, in one embodiment, theinfant is administered a days 1-2 formula having an energy content offrom about 200 kcal/L to about 360 kcal/L during the first two daysfollowing birth, and is subsequently administered a days 3-9 formulahaving an energy content of from about 360 kcal/L to less than 600kcal/L on days 3 to 9 following birth. Optionally, the days 3-9 formulamay be administered past day 9 following birth, or alternatively, ahigher calorie formula (Including full calorie formulas) may beadministered starting on day 10 following birth. The formula (s)administered to the infant will typically be administered daily atintake volumes as described hereinbefore.

In another aspect, the present disclosure is directed to a method ofimproving protein absorption in an infant comprising administering tothe infant any of the low micronutrient infant formulas of the presentdisclosure. Preferably, the infant is a newborn infant. The lowmicronutrient infant formula may be any of those set forth above.

These methods may also further comprise administering two or moredifferent infant formulas to the infant. For instance, in oneembodiment, the infant is administered a low calorie infant formula(having either a high or low micronutrient content) having an energycontent of from about 200 kcal/L to about 360 kcal/L (e.g., a days 1-2formula), during the first two days following birth. The infant may thensubsequently be administered a low calorie infant formula (having eithera high or low micronutrient content) that has an energy content of fromabout 360 kcal/L to less than 600 kcal/L (e.g., a days 3-9 formula) ondays 3 to 9 following birth. Optionally, the days 3-9 formula may beadministered past day 9 following birth, or alternatively, a highercalorie formula (Including full calorie formulas) may be administeredstarting on day 10 following birth. In embodiments where the low calorieinfant formulas have a low micronutrient content, the amounts ofmicronutrients included in the formulas may be any of those set forthabove. The formula (s) administered to the infant will typically beadministered daily at intake volumes as described hereinbefore.

Tolerance

The present disclosure is also directed to a method of improving theinfant formula tolerance of an infant. Infant formula intolerance is anon-immune system associated reaction that may be evidenced by behavioror by stool or feeding pattern changes, such as increased spit-up orvomiting, an increased number of stools, more watery stools, blackstools, and increased fussiness. Infant formula intolerance is mostoften associated with gastrointestinal symptoms (e.g., stool patterns,gas, spit-up) as well as behavior characteristics (e.g., acceptance offormula, fussing and crying). Infants suffering from formula intolerancemay also experience gastroesophageal reflux.

It has now unexpectedly been discovered that infants have a greatertolerance for an infant formula having a low energy content than forfull calorie formulas. Specifically, it has been discovered that lowcalorie infant formulas demonstrate a faster rate of protein hydrolysisand digestion, produce less Maillard reaction products (which cannot bebroken down and absorbed) upon consumption, and have a faster rate ofgastric emptying than do full calorie formulas. The faster gastricemptying leads to decreased gastroesophageal reflux, and improvedtolerance of the formula.

The low calorie infant formulas of the present disclosure may thus beused to decrease the incidence of gas, and/or spit up in infants. Thelow calorie infant formulas of the present disclosure may also be usedto increase the rate of gastric emptying in the infant and reduce thedegree of Maillard reaction products resulting from formula consumption,as compared to full calorie infant formulas.

The low calorie infant formulas can be administered to any infant,preterm or full term, and especially any infant that can benefit fromreceiving an infant formula having a low energy content that also hashigh tolerance. In some embodiments, the low calorie infant formulas ofthe present disclosure are administered to newborn infants.

Thus, in one aspect, the present disclosure is directed to a method ofimproving the infant formula tolerance of an infant. The methodcomprises identifying an infant having infant formula intolerance andadministering to the infant any one or more of the low calorie infantformulas of the present disclosure. Infants having infant formulaintolerance can include infants having any one or more of the symptomsof formula intolerance. Such symptoms include, but are not limited to,stool or feeding pattern changes, such as increased spit-up or vomiting,an increased number of stools, more watery stools, black stools,increased fussiness, crying, gas, and lack of acceptance of formula.Upon administration of a low calorie infant formula of the presentdisclosure, some or all of the symptoms of formula intolerance may bereduced or eliminated.

As noted above, any of the low calorie infant formulas of the presentdisclosure may be used in this method. The low calorie infant formulamay have a low micronutrient content, or, in some embodiments, may havea high micronutrient content, and may be a days 1-2 or a days 3-9formula. In one embodiment, the low calorie infant formula has an energycontent of from about 200 to about 600 kilocalories per liter offormula.

The method may also further comprise administering two or more differentinfant formulas to the infant. For instance, in one embodiment, theinfant is administered a days 1-2 formula having an energy content offrom about 200 kcal/L to about 360 kcal/L during the first two daysfollowing birth, and is subsequently administered a days 3-9 formulahaving an energy content of from about 360 kcal/L to less than 600kcal/L on days 3 to 9 following birth. Optionally, the days 3-9 formulamay be administered past day 9 following birth, or alternatively, ahigher calorie formula (Including full calorie formulas) may beadministered starting on day 10 following birth. The formula (s)administered to the infant will typically be administered daily atintake volumes as described hereinbefore.

In another aspect, the present disclosure is directed to a method forimproving the infant formula tolerance of an infant comprisingadministering to the infant any of the low micronutrient infant formulasof the present disclosure. Preferably, the infant is a newborn infant.The low micronutrient infant formula comprises micronutrients and atleast one macronutrient selected from the group consisting of protein,carbohydrate, fat, and combinations thereof. In one embodiment, the lowmicronutrient infant formula has an energy content of from about 200kcal/L to less than 600 kcal/L, wherein at least 65% of themicronutrients are included in the infant formula in an amount that isfrom about 30% to about 80% of conventional amounts of correspondingmicronutrients, on a per volume basis. In another embodiment, the lowmicronutrient infant formula has an energy content of from about 200kcal/L to about 360 kcal/L, wherein at least 45% of the micronutrientsare included in the infant formula in an amount that is from about 30%to about 65% of conventional amounts of corresponding micronutrients, ona per volume basis. In still another embodiment, the low micronutrientinfant formula has an energy content of from about 360 kcal/L to lessthan 600 kcal/L, wherein at least 30% of the micronutrients are includedin the infant formula in an amount that is from about 55% to about 80%of conventional amounts of corresponding micronutrients, on a per volumebasis. The low calorie infant formula may be a days 1-2 or a days 3-9formula.

These methods may also further comprise administering two or moredifferent infant formulas to the infant. For instance, in oneembodiment, the infant is administered a low calorie infant formula(having either a high or low micronutrient content) having an energycontent of from about 200 kcal/L to about 360 kcal/L (e.g., a days 1-2formula), during the first two days following birth. The infant may thensubsequently be administered a low calorie infant formula (having eithera high or low micronutrient content) that has an energy content of fromabout 360 kcal/L to less than 600 kcal/L (e.g., a days 3-9 formula) ondays 3 to 9 following birth. Optionally, the days 3-9 formula may beadministered past day 9 following birth, or alternatively, a highercalorie formula (Including full calorie formulas) may be administeredstarting on day 10 following birth. In embodiments where the low calorieinfant formulas have a low micronutrient content, the amounts ofmicronutrients included in the formulas may be any of those set forthabove. The formula (s) administered to the infant will typically beadministered daily at intake volumes as described hereinbefore.

In still another embodiment, the present disclosure is directed to amethod for inhibiting gastroesophageal reflux in an infant. The methodcomprises identifying an infant having gastroesophageal reflux, andadministering to the infant any one or more of the low calorie infantformulas of the present disclosure. Preferably, the infant is a newborninfant.

Gastroesophageal reflux (GER) occurs when stomach contents reflux intothe esophagus and out of the mouth, resulting in regurgitation, spittingup, and/or vomiting. Symptoms of GER include spitting up, vomiting,coughing, irritability, poor feeding, bloody stool, and combinationsthereof GER may also occur when infants cough, cry, or strain. Forpurposes of the present disclosure, the term “inhibitinggastroesophageal reflux” is intended to include treating, preventing,and/or decreasing the rate of occurrence of GER and/or at least one ofits symptoms. Without wishing to be bound to any particular theory, itis believed that the low calorie infant formula of the presentdisclosure has a faster rate of gastric emptying (i.e., the rate atwhich contents pass through the stomach), which leads to decreasedgastroesophageal reflux, as compared to full calorie formulas.

As noted above, any of the low calorie infant formulas of the presentdisclosure may be used in this method. The low calorie infant formulamay have a low micronutrient content, or, in some embodiments, may havea high micronutrient content, and may be a days 1-2 or a days 3-9formula. In one embodiment, the infant formula has an energy content offrom about 200 kcal/L to less than 600 kcal/L of formula.

The method may also further comprise administering two or more differentinfant formulas to the infant. For instance, in one embodiment, theinfant is administered a days 1-2 formula having an energy content offrom about 200 kcal/L to about 360 kcal/L during the first two daysfollowing birth, and is subsequently administered a days 3-9 formulahaving an energy content of from about 360 kcal/L to less than 600kcal/L on days 3 to 9 following birth. Optionally, the days 3-9 formulamay be administered past day 9 following birth, or alternatively, ahigher calorie formula (including full calorie formulas) may beadministered starting on day 10 following birth. The formula (s)administered to the infant will typically be administered daily atintake volumes as described hereinbefore.

In another aspect, the present disclosure is directed to a method forinhibiting gastroesophageal reflux in an infant comprising administeringto the infant any one or more of the low micronutrient infant formulasof the present disclosure. Preferably, the infant is a newborn infant.The low micronutrient infant formula may be any of those set forthabove.

These methods may also further comprise administering two or moredifferent infant formulas to the infant. For instance, in oneembodiment, the infant is administered a low calorie infant formula(having either a high or low micronutrient content) having an energycontent of from about 200 kcal/L to about 360 kcal/L (e.g., a days 1-2formula), during the first two days following birth. The infant may thensubsequently be administered a low calorie infant formula (having eithera high or low micronutrient content) that has an energy content of fromabout 360 kcal/L to less than 600 kcal/L (e.g., a days 3-9 formula) ondays 3 to 9 following birth. Optionally, the days 3-9 formula may beadministered past day 9 following birth, or alternatively, a highercalorie formula (Including full calorie formulas) may be administeredstarting on day 10 following birth. In embodiments where the low calorieinfant formulas have a low micronutrient content, the amounts ofmicronutrients included in the formulas may be any of those set forthabove. The formula (s) administered to the infant will typically beadministered daily at intake volumes as described hereinbefore.

In another aspect, the present disclosure is directed to a method forincreasing the rate of gastric emptying in an infant comprisingadministering to the infant any one or more of the low micronutrientinfant formulas of the present disclosure. Preferably, the infant is anewborn infant. The low micronutrient infant formula may be any of thoseset forth above.

These methods may also further comprise administering two or moredifferent infant formulas to the infant. For instance, in oneembodiment, the infant is administered a low calorie infant formula(having either a high or low micronutrient content) that has an energycontent of from about 200 kcal/L to about 360 kcal/L (e.g., a days 1-2formula), during the first two days following birth. The infant may thensubsequently be administered a low calorie infant formula (having eithera high or low micronutrient content) that has an energy content of fromabout 360 kcal/L to less than 600 kcal/L (e.g., a days 3-9 formula) ondays 3 to 9 following birth. Optionally, the days 3-9 formula may beadministered past day 9 following birth, or alternatively, a highercalorie formula (Including full calorie formulas) may be administeredstarting on day 10 following birth. The amounts of micronutrientsincluded in the formulas may be any of those set forth above. Theformula (s) administered to the infant will typically be administereddaily at intake volumes as described hereinbefore.

Kits

The present disclosure further provides kits comprising two or more ofthe low calorie infant formulas of the present disclosure.

For instance, in some embodiments, the kit may comprise at least onedays 1-2 formula and at least one days 3-9 formula. Preferably, the kitwill comprise sufficient amounts of the days 1-2 formula to provide aninfant with adequate nutrition during the first two days followingbirth, and sufficient amounts of the days 3-9 formula to provide aninfant with adequate nutrition for at least days 3-9 following birth.The infant formulas included in the kit may be in any suitable form,including, for example, a ready-to-feed liquid, a concentrated liquid, apowder, or combinations thereof. The kit may include low calorie, lowmicronutrient formulas and/or low calorie, high micronutrient formulas.

Optionally, the kits may further comprise instructions for use of thekit. For instance, the instructions may describe how to use theformulas, e.g., may indicate that the days 1-2 formulas should beadministered on the first two days following birth and that the days 3-9formulas should be administered on days 3-9 following birth; maydescribe a daily administration schedule for the formulas; and/or maydescribe how to practice any of the methods described in the presentdisclosure. The instructions may further optionally describe how toreconstitute any powder infant formulas included in the kit.

In addition to the infant formulas and optional instructions, the kitcan also include additional components, such as one or more baby bottlesof various sizes, one or more baby bottle liners of various sizes, babybottle nipples, and the like.

EXAMPLES

The following examples illustrate specific embodiments and/or featuresof the infant formulas and methods of the present disclosure. Theexamples are given solely for the purpose of illustration and are not tobe construed as limitations of the present disclosure, as manyvariations thereof are possible without departing from the spirit andscope of the disclosure. All exemplified amounts are weight percentagesbased upon the total weight of the composition, unless otherwisespecified.

Unless otherwise specified, the retort sterilized and aseptic sterilizedformulas prepared in accordance with the manufacturing methods describedherein, are ready-to-feed liquid formulas.

Examples 1-8

In these examples, 2 oz. retort sterilized days 1-2 and days 3-9 infantformulas were prepared with either high or low micronutrient content.The ingredients used to prepare the formulas are set forth in Tables 1and 2 below.

TABLE 1 Days 1-2 Formulas Formula 1 Formula 2 Formula 3 Formula 4 Units(days 1-2) (days 1-2) (days 1-2) (days 1-2) Energy Kcal/L 270 270 250250 Micronutrient content low low high high Ingredients (Amount Per 1000Kg batch) Water kg Q.S. Q.S. Q.S. Q.S. Lactose kg 23.2 23.1 15.5 15.2Nonfat Dry Milk kg 11.0 11.0 11.0 11.3 Galactooligosaccharides kg 4.404.40 4.40 4.40 High Oleic Safflower Oil kg 5.34 5.35 5.33 5.37 Soy Oilkg 4.00 4.00 3.99 4.00 Coconut Oil kg 3.82 3.82 3.81 3.84 Whey Proteinkg 2.70 2.70 2.70 2.86 Concentrate 1N KOH g 1340 1.40 1340 1340 Potassium Hydroxide g 67.0 70.0 67.0 67.0 Calcium Phosphate g 327.1249.8 1090 770.2 Dibasic Potassium Citrate g 3.10 1.24 1370 1240 CalciumCitrate g 351.0 578.8 752.6 768.9 Ascorbic Acid g 727.5 727.5 727.5727.5 ARA Oil g 367.9 367.9 367.9 367.9 Nucleotide-Choline g 328.5 328.5328.5 328.5 Premix Dicalcium Phosphate g — — — — Magnesium Chloride g16.8 102.6 460.9 450.7 Sodium Chloride g 45.7 28.5 325.8 186.7 SoyLecithin g 143.0 143.0 143.0 143.0 Distilled Monoglycerides g 143.0143.0 143.0 143.0 Vitamin/Mineral/Taurine Premix g 31.4 57.1 157.0 157.0 Taurine g 9.60 17.5 48.0 48.0  m-Inositol g 6.97 12.7 34.85 34.85  ZincSulfate g 3.21 5.85 16.07 16.07  Niacinamide g 2.05 3.73 10.24 10.24 Calcium Pantothenate g 1.23 2.23 6.14 6.14  Ferrous Sulfate g 1.07 1.955.37 5.37  Cupric Sulfate mg 377 686 1890 1890  Thiamine Chloride mg 318578 1590 1590  HCL  Riboflavin mg 140 255 701 701  Pyridoxine HCL mg 128234 642 642  Folic Acid mg 43.2 78.5 216 216  Manganese Sulfate mg 36.666.5 183 183  Biotin mg 12.4 22.6 62.0 62.0  Sodium Selenate mg 7.4413.5 37 37  Cyanocobalamin mg 0.990 1.8 4.95 4.95 DHA Oil g 137.9 137.9137.9 137.9 Potassium Chloride g 46.3 52.4 As needed 60.7 CholineChloride g 58.9 21.5 88.9 54.0 Ferrous Sulfate g 5.80 23.20 60.9 60.9Carrageenan g 175.0 175.0 175.0 175.0 Vitamin A, D3, E, K1 g 22.8 19.047.5 47.5  RRR α-Tocopherol Acetate g 4.61 3.84 9.6 9.6  Vitamin APalmitate mg 867 721.5 1800 1800  Vitamin K1 mg 50.2 41.8 104.5 104.5 Vitamin D3 mg 6.08 5.06 12.65 12.65 Citric Acid g 29.8 29.8 29.8 29.8Mixed Carotenoid Premix g 23.8 23.8 23.8 23.8  Lycopene mg 119 119 119119  Lutein mg 50 50 50 50  Beta-carotene mg 26.2 26.2 26.2 26.2Inositol g 33.1 6.6 12.9 12.9 L-Carnitine g 6.38 1.31 6.38 3.28Riboflavin mg — 466.0 882 882

TABLE 2 Days 3-9 Formulas Formula 5 Formula 6 Formula 7 Formula 8 Units(days 3-9) (days 3-9) (days 3-9) (days 3-9) Energy Kcal/L 406 406 406410 Micronutrient content low low low high Ingredients (Amount Per 1000Kg Batch) Water kg Q.S. Q.S. Q.S. Q.S. Lactose kg 37.0 37.2 37.5 35.50Nonfat Dry Milk kg 16.3 16.2 16.2 16.30 Galactooligosaccharides kg 8.638.63 8.63 8.63 High Oleic Safflower Oil kg 7.72 7.72 7.72 7.72 Soy Oilkg 5.78 5.78 5.78 5.78 Coconut Oil kg 5.52 5.52 5.52 5.51 Whey ProteinConcentrate kg 4.00 4.00 4.00 4.00 1N KOH kg 1.34 1.34 0.8035 1.34 Potassium Hydroxide g 67.0 67.0 40.2 67.0 Calcium Phosphate kg 0.309 —— — Dibasic Potassium Citrate kg 0.00186 0.00186 0.00186 1.06 CalciumCitrate g 687.6 583.5 583.5 261.1 Ascorbic Acid g 727.5 727.5 436.5727.5 ARA Oil g 378.2 378.2 378.2 378.2 Nucleotide-Choline g 319.7 319.7319.7 319.7 Premix Ultra Micronized g — 226.8 226.8 1470 TricalciumPhosphate Magnesium Chloride g 122.5 147.7 147.7 288.1 Sodium Chloride g— — 235.8 Soy Lecithin g 206.0 206.0 206.0 206.0 DistilledMonoglycerides g 206.0 206.0 206.0 206.0 Vitamin/Mineral/Taurine Premixg 85.6 115.7 115.7 142.7  Taurine g 26.2 35.4 35.4 43.6  m-Inositol g19.0 25.7 25.7 31.7  Zinc Sulfate g 8.76 11.8 11.8 14.61  Niacinamide g5.59 7.55 7.55 9.31  Calcium Pantothenate g 3.35 4.53 4.53 5.58  FerrousSulfate g 2.93 3.96 3.96 4.88  Cupric Sulfate g 1.03 1.39 1.39 1.71 Thiamine Chloride g 0.8667 1.17 1.17 1.44  HCL  Riboflavin mg 382.2516.6 516.6 637  Pyridoxine HCl mg 350.1 473.2 473.2 584  Folic Acid mg117.7 159.1 159.1 196  Manganese Sulfate mg 99.7 134.7 134.7 166  Biotinmg 33.8 45.7 45.7 56.0  Sodium Selenate mg 20.3 27.4 27.4 34 Cyanocobalamin mg 2.7 3.64 3.64 4.5 DHA Oil g 137.9 137.9 137.9 137.9Potassium Chloride g 108.7 111.3 111.3 129.5 Choline Chloride g 32.432.4 32.4 88.9 Ferrous Sulfate g 34.8 37.5 37.5 60.9 Carrageenan g 175.0175.0 175.0 175.0 Vitamin A, D3, E, K1 g 28.5 30.2 30.2 44.8  RRRα-Tocopherol g 5.8 6.11 6.11 9.1  Acetate  Vitamin A Palmitate g 1.081.15 1.15 1.7  Vitamin K1 mg 62.7 66.4 66.4 98.5  Vitamin D3 mg 7.6 8.048.04 11.9 Citric Acid g 29.8 29.8 29.8 29.8 Mixed Carotenoid Premix g23.8 23.8 23.8 23.8  Lycopene mg 119 119 119 119  Lutein mg 50 50 50 50 Beta-carotene mg 26.2 26.2 26.2 26.2 Inositol g — — — 12.9 L-Carnitineg 1.97 2.31 2.31 5.51 Riboflavin g 0.70 0.699 0.699 1.50 Vitamin A mg —770 770 780  Vitamin A Palmitate mg — 420 420 425 Copper Sulfate mg — —— 391

The formulas were prepared by making at least two separate slurries thatwere later blended together, heat treated, standardized, and terminallysterilized. Initially, a carbohydrate-mineral slurry was prepared bydissolving the selected carbohydrates (e.g. lactose,galactooligosacchardies) in water at 74-79° C., followed by the additionof citric acid, magnesium chloride, potassium chloride, potassiumcitrate, choline chloride, and sodium chloride. The resulting slurry washeld under moderate agitation at 49-60° C. until it was later blendedwith the other prepared slurries.

A protein-in-oil slurry was prepared by combining the high oleicsafflower oil, coconut oil, monoglycerides, and soy lecithin underagitation and heating to 66-79° C. Following a 10-15 minute hold time,soybean oil, oil soluble vitamin premix, mixed carotenoid premix,carrageenan, vitamin A, calcium citrate, dicalcium phosphate, ARA oil,DHA oil, and whey protein concentrate were then added to the slurry. Theresulting oil slurry was held under moderate agitation at 49-60° C.until it was later blended with the other prepared slurries.

Water was heated to 49-60° C. and then combined with thecarbohydrate-mineral slurry, nonfat milk, and the protein-in-oil slurryunder adequate agitation. The pH of the resulting blend was adjustedwith potassium hydroxide. This blend was held under moderate agitationat 49-60° C.

The resulting blend was heated to 74-79° C., emulsified through a singlestage homogenizer to 900-1100 psig, and then heated to 144-147° C., forabout 5 seconds. The heated blend was passed through a flash cooler toreduce the temperature to 88-93° C. and then through a plate cooler tofurther reduce the temperature to 74-85° C. The cooled blend was thenhomogenized at 2900-3100/400-600 psig, held at 74-85° C. for 16 seconds,and then cooled to 2-7° C. Samples were taken for analytical testing.The mixture was held under agitation at 2-7° C.

A water-soluble vitamin (WSV) solution and an ascorbic acid solutionwere prepared separately and added to the processed blended slurry. Thevitamin solution was prepared by adding the following ingredients towater with agitation: potassium citrate, ferrous sulfate, WSV premix,L-carnitine, copper sulfate, riboflavin, inositol, and thenucleotide-choline premix. The ascorbic acid solution was prepared byadding potassium hydroxide and ascorbic acid to a sufficient amount ofwater to dissolve the ingredients. The ascorbic acid solution pH wasthen adjusted to 5-9 with potassium hydroxide.

The blend pH was adjusted to a specified pH range of 7.1-7.6 withpotassium hydroxide (varied by product) to achieve optimal productstability. The completed product was then filled into suitablecontainers and terminally sterilized.

Examples 9-11

In these examples, 32 oz. aseptic sterilized days 3-9 infant formulaswere prepared with either high or low micronutrient content. Theingredients used to prepare the formulas are set forth in Table 3 below.

TABLE 3 Formula 9 Formula 10 Formula 11 Units (days 3-9) (days 3-9)(days 3-9) Energy Kcal/L 406 410 410 Micronutrient Content low high highIngredients Amount per 1000 kg batch Water kg Q.S. Q.S. Q.S. Lactose kg37.0 33.7 34.03 Nonfat Dry Milk kg 16.3 17.0 16.47Galactooligosaccharides kg 8.63 8.63 8.63 High Oleic Safflower Oil kg7.72 7.83 7.72 Soy Oil kg 5.78 5.87 5.78 Coconut Oil kg 5.52 5.60 5.51Whey Protein Concentrate kg 4.00 4.19 4.05 1N KOH kg 1.85 1.85 1.85 Potassium Hydroxide g 92.5 92.5 92.5 Calcium Citrate g 675.0 716.8993.9 Calcium Phosphate Dibasic g 577.4 1170 1390 Ascorbic Acid g 431.7431.7 431.7 ARA Oil g 378.2 378.2 378.2 Nucleotide-Choline Premix g319.7 319.7 319.7 Soy Lecithin g 206.0 206.0 206.0 DistilledMonoglycerides g 206.0 206.0 206.0 Carrageenan g 200.0 240.0 200.0 DHAOil g 137.9 137.9 137.9 Magnesium Chloride g 128.9 279.3 285.9 PotassiumChloride g 118.5 213.9 122.4 Choline Chloride g 88.9 54.0 88.9Vitamin/Mineral/Taurine Premix g 41.4 142.7 142.7  Taurine g 12.7 43.643.6  m-Inositol g 9.19 31.7 31.7  Zinc Sulfate g 4.24 14.61 14.61 Niacinamide g 2.70 9.31 9.31  Calcium Pantothenate g 1.62 5.58 5.58 Ferrous Sulfate g 1.42 4.88 4.88  Cupric Sulfate mg 497 1710 1710 Thiamine Chloride HCl mg 419 1440 1440  Riboflavin mg 185 637 637 Pyridoxine HCl mg 169 584 584  Folic Acid mg 56.9 196 196  ManganeseSulfate mg 48.2 166 166  Biotin mg 16.4 56.0 56.0  Sodium Selenate mg9.81 34 34  Cyanocobalamin mg 1.3 4.5 4.5 Sodium Chloride g 32.1 65.4231.9 Vitamin A, D3, E, K1 g 30.9 44.8 44.8  RRR Alpha-TocopherylAcetate g 6.24 9.1 9.1  Vitamin A Palmitate g 1.17 1.7 1.7  Vitamin K1mg 67.9 98.5 98.5  Vitamin D3 mg 8.22 11.9 11.9 Citric Acid g 29.8 29.829.8 Inositol g 25.8 12.9 12.9 Mixed Carotenoid Premix g 23.8 23.8 23.8 Lycopene mg 119 119 119  Lutein mg 50 50 50  Beta-Carotene mg 26.2 26.226.2 Ferrous Sulfate g 16.2 60.9 60.9 L-Carnitine g 5.51 3.28 5.51Potassium Citrate g 3.10 895.0 1060 Riboflavin mg 599 1500 1500 VitaminA mg — 780 780  Vitamin A Palmitate mg — 425 425 Copper Sulfate mg — —391

The formulas were prepared by making at least two separate slurries thatwere later blended together, heat treated, standardized, and thenaseptically processed and filled. Initially, a carbohydrate-mineralslurry was prepared by dissolving the selected carbohydrates (e.g.lactose, galactooligosacchardies) in water at 74-79° C., followed by theaddition of citric acid, magnesium chloride, potassium chloride,potassium citrate, choline chloride, and sodium chloride (mineralsvaried by formulation). The resulting slurry was held under moderateagitation at 49-60° C. until it was later blended with the otherprepared slurries.

A protein-in-oil slurry was prepared by combining high oleic saffloweroil, coconut oil, monoglycerides, and soy lecithin under agitation andheating to 66-79° C. Following a 10-15 minute hold time, soybean oil,oil soluble vitamin premix, mixed carotenoid premix, carrageenan,calcium citrate, calcium phosphate dibasic, ARA oil, DHA oil, and wheyprotein concentrate were added to the slurry. The resulting oil slurrywas held under moderate agitation at 49-60° C. until it was laterblended with the other prepared slurries.

Water was heated to 49-60° C. and then combined with thecarbohydrate-mineral slurry, nonfat milk, and the protein-in-oil slurryunder adequate agitation. The pH of the resulting blend was adjustedwith potassium hydroxide. This blend was held under moderate agitationat 49-60° C.

The resulting blend was heated to 74-79° C., emulsified through a singlestage homogenizer to 900-1100 psig, and then heated to 144-147° C., forabout 5 seconds. The heated blend was passed through a flash cooler toreduce the temperature to 88-93° C., and then through a plate cooler tofurther reduce the temperature to 74-85° C. The cooled blend was thenhomogenized at 2900-3100/400-600 psig, held at 74-85° C. for 16 seconds,and then cooled to 2-7° C. Samples were taken for analytical testing.The mixture was held under agitation at 2-7° C.

A water-soluble vitamin (WSV) solution and an ascorbic acid solutionwere prepared separately and added to the processed blended slurry. Thevitamin solution was prepared by adding the following ingredients towater with agitation: potassium citrate, ferrous sulfate, WSV premix,L-carnitine, riboflavin, inositol, and the nucleotide-choline premix.The ascorbic acid solution was prepared by adding potassium hydroxideand ascorbic acid to a sufficient amount of water to dissolve theingredients. The ascorbic acid solution pH was then adjusted to 5-9 withpotassium hydroxide.

The blend pH was adjusted to a pH range of 6.8-7.0 with potassiumhydroxide to achieve optimal product stability. The standardized blendthen received a second heat treatment through an aseptic processor. Theblend was preheated to 63-74° C. and homogenized at 200 psig. The blendwas further heated to 141-144° C. and passed through a hold tube. Theheated blend was cooled to reduce the temperature to 74-85° C., and thenhomogenized at 1200/200 psig. The blend was further cooled to 16-27° C.,and then aseptically filled into suitable containers at 21° C.

Examples 12-15

In these examples, powder days 1-2 and days 3-9 infant formulas wereprepared with either low or high micronutrient content. The ingredientsused to prepare the formulas are set forth in Table 4 below.

TABLE 4 Formula 12 Formula 13 Formula 14 Formula 15 (days 1-2) (days1-2) (days 3-9) (days 3-9) Kcal/L 270 250 406 420 Nutrient Content lowhigh low high Ingredients Units Amount per 1000 kg batch Lactose kg376.90 288.6 406.4 380.4 Non-Fat Dry Milk kg 223.00 223.1 201.1 201.1High Oleic Safflower Oil kg 109.30 108.5 97.69 97.7Galactooligosaccharides kg 81.70 84.7 104.1 104.10 Soy Oil kg 81.70 82.474.21 74.2 Coconut Oil kg 75.30 75.9 68.36 68.4 Whey Protein Concentratekg 48.80 54.9 49.50 49.5 Potassium Citrate kg 8.52 42.6 11.12 22.0 ARAOil kg 7.20 7.43 4.643 4.57 Whey Protein Hydrolysate kg 6.80 — — —Calcium Carbonate kg 3.76 — 2.839 1.5 Tricalcium Phosphate kg — 24.12.638 10.9 DHA Oil kg 2.70 2.8 1.752 1.7 Ascorbic Acid kg 2.03 3.202.006 2.0 Nucleotide-Choline Premix kg 2.01 5.9 2.346 3.6 PotassiumChloride kg 1.154 — 1.219 — Vitamin/Mineral/Taurine kg 1.116 2.8 1.1161.7 Premix  Taurine g 341 859.9 341 528.9  m-Inositol g 248 624.3 248384.0  Zinc Sulfate g 114 287.9 114 177.1  Niacinamide g 72.8 183.5 72.8112.9  Calcium Pantothenate g 43.7 110 43.7 67.7  Ferrous Sulfate g 38.296.3 38.2 59.2  Cupric Sulfate g 13.4 33.8 13.4 20.8  Thiamine ChlorideHCl g 11.3 28.5 11.3 17.5  Riboflavin g 4.98 12.60 4.98 7.72  PyridoxineHCl g 4.58 11.5 4.58 7.07  Folic Acid g 1.53 3.9 1.53 2.4  ManganeseSulfate g 1.3 3.27 1.3 2.01  Biotin mg 441 1100 441 683  Sodium Selenatemg 264 666.1 264 410  Cyanocobalamin mg 35.1 88.6 35.1 54.5 Soy Lecithinkg 1.120 1.1 1.112 1.1 Magnesium Chloride kg 0.839 6.6 1.437 3.4Potassium Chloride kg — 2.6 — 2.3 Ascorbyl Palmitate g 459.25 348.1313.5 313.6 Carotenoid Premix g 454.02 463.0 286.6 286.6  Lycopene g2.27 2.27 1.43 1.41  Lutein mg 953 953 602 589.9  Beta-Carotene mg 499499 315 308.9 Ferrous Sulfate g 453.5 1100 453.6 703.1 Choline Chlorideg 432.1 1100 432.1 670.2 Sodium Chloride g 388.0 7100 1138 2900 VitaminA, D3, E, K1 g 385.24 914.5 327.3 568.8  RRR α-Tocopheryl g 77.9 184.966.2 115.0  Acetate  Vitamin A Palmitate g 14.63 34.7 12.4 21.6  VitaminK1 mg 847 2000 720 1250  Vitamin D3 mg 102.3 243.5 87.1 151.4 MixedTocopherols g 246.3 153.4 138.2 138.2 L-Carnitine g 26.3 66.3 23.3 40.8Riboflavin g 3.2 8.0 3.2 4.9 1N Potassium Hydroxide as needed as neededas needed as needed

The formulas were prepared by making at least two separate slurries thatwere later blended together, heat treated, standardized, heat treated asecond time, evaporated to remove water, and finally spray dried.Initially, a carbohydrate-mineral slurry was prepared by dissolving theselected carbohydrates (e.g. lactose, galactooligosaccharides) in waterat 60-71° C., followed by the addition of magnesium chloride, potassiumchloride, potassium citrate, choline chloride, and sodium chloride(minerals vary depending on formulation). The resulting slurry was heldunder moderate agitation at 49-60° C. until it was later blended withthe other prepared slurries.

A protein-in-oil slurry was prepared by combining high oleic saffloweroil, soybean oil, and coconut oil at 49-60° C., followed by the additionof ascorbyl palmitate, mixed tocopherols, soy lecithin, oil solublevitamin premix, whey protein concentrate, whey protein hydrolysate (insome cases), carotenoid premix, and calcium carbonate (and/or tricalciumphosphate). The resulting oil slurry was held under moderate agitationat 38-49° C. until it was later blended with the other preparedslurries.

Water, the carbohydrate-mineral slurry, non fat milk, and theprotein-in-oil slurry, were combined under adequate agitation. The pH ofthe resulting blend was adjusted with potassium hydroxide. This blendwas held under moderate agitation at 49-60° C. The ARA and DHA oil wereadded following the pH adjustment and prior to processing.

The resulting blend was heated to 71-77° C., emulsified through a singlestage homogenizer to a maximum of 300 psig, and then heated to 82-88°C., for about 5 seconds. The heated blend was passed through a flashcooler to reduce the temperature to 77-82° C. and then through a platecooler to further reduce the temperature to 71-77° C. The cooled blendwas then homogenized at 2400-2600/400-600 psig, held at 74-85° C. for 16seconds, and then cooled to 2-7° C. Samples were taken for analyticaltesting. The mixture was held under agitation at 2-7° C.

A water-soluble vitamin (WSV) solution and an ascorbic acid solutionwere prepared separately and added to the processed blended slurry. Thevitamin solution was prepared by adding the following ingredients towater with agitation: potassium citrate, ferrous sulfate, WSV premix,L-carnitine, riboflavin, and the nucleotide-choline premix (specificingredients vary by formulation). The ascorbic acid solution wasprepared by adding potassium hydroxide and ascorbic acid to a sufficientamount of water to dissolve the ingredients. The ascorbic acid solutionpH was then adjusted to 5-9 with potassium hydroxide.

The blend pH was adjusted to a pH range of 6.60-6.90 with potassiumhydroxide to achieve optimal product stability. The standardized blendthen received a second heat treatment. The blend was originally heatedto 66-82° C., and then further heated to 118-124° C. for about 5seconds. The heated blend was then passed through a flash cooler toreduce the temperature to 71-82° C. Following heat treatment, the blendwas evaporated down to a density of 1.15-1.17 g/mL.

The evaporated blend was passed through a spray drier, targeting amoisture level of 2.5% in the finished powder. The finished powder thenunderwent agglomeration with water as the binder solution. The completedproduct was then packaged into suitable containers.

Example 16

In this example, the effect of energy content on the buffering capacityand buffering strength of infant formula was evaluated. Specifically,the buffering capacity and buffering strength of various days 1-2 anddays 3-9 infant formulas of the present disclosure were determined andcompared to the buffering capacity and buffering strength of acommercially available powder control infant formula, a commerciallyavailable ready-to-feed 2 oz. retort sterilized control infant formula,a commercially available ready-to-feed 32 oz. aseptic sterilized controlinfant formula, and human milk. The ingredients used to prepare thecontrol formulas are set forth in Table 5 below.

TABLE 5 Control Control Control Formula 1 Formula 2 Formula 3 (powder)(retort) (aseptic) Kcal/L 676 676 676 Ingredients Units Amount per 1000kg batch Water kg — Q.S. Q.S. Condensed Skim Milk kg 698.5 83.61 86.64Lactose kg 386.0 54.88 54.7 High Oleic Safflower Oil kg 114.4 14.07 14.0Soy Oil kg 85.51 10.54 10.5 Coconut Oil kg 78.76 10.05 10.0Galactooligosaccharides kg 69.50 8.630 8.60 Whey Protein Concentrate kg51.08 6.120 6.52 Potassium Citrate g 9168 518.3 418.07 Calcium Carbonateg 4054 508.5 477.16 ARA Oil g 2949 355.6 378.16 Nucleotide-CholinePremix g 2347 293.2 293.26 Potassium Chloride g 1295 208.5 282.24Carrageenan g — 175.0 240.00 Ascorbic Acid G 1275 727.5 582.12 SoyLecithin G 1120 534.6 356.11 Stabilizer G — 534.6 356.11Vitamin/Mineral/Taurine Premix G 1116 142.8 142.77  Taurine G 340.543.66 43.654  m-Inositol G 247.9 31.70 31.695  Zinc Sulfate G 114.214.62 14.617  Niacinamide G 72.78 9.323 9.3157  Calcium Pantothenate G44.16 5.587 5.5860  Ferrous Sulfate G 39.24 4.880 4.8870  Cupric SulfateG 13.68 1.714 1.7143  Thiamine Chloride HCl G 11.30 1.445 1.4456 Riboflavin Mg 4985 637.6 637.47  Pyridoxine HCl Mg 4572 584.1 583.96 Folic Acid Mg 1535 196.4 215.72  Manganese Sulfate Mg 1306 166.3 166.25 Biotin Mg 441.0 56.41 56.390  Sodium Selenate Mg 261.8 33.82 33.820 Cyanocobalamin Mg 35.17 4.493 4.500 DHA Oil G 1113 135.4 130.01Magnesium Chloride G 1038 141.5 140.46 Sodium Chloride G 579.4 as neededas needed Ferrous Sulfate G 453.6 58.02 58.03 Choline Chloride G 432.154.02 50.02 Vitamin A, D3, E, K1 G 377.2 47.50 44.76  RRRAlpha-Tocopheryl Acetate G 76.23 9.604 9.0507  Vitamin A Palmitate G14.32 1.803 1.6998  Vitamin K1 Mg 829.3 104.5 98.47  Vitamin D3 Mg 100.412.65 11.92 Citric Acid G — 29.80 29.77 Ascorbyl Palmitate G 361.3 — —Carotenoid Premix G 350.1 23.80 42.91  Lycopene Mg 1720 119.0 214.55 Lutein Mg 735 49.98 90.11  Beta-Carotene Mg 385 26.18 47.201 MixedTocopherols G 159.2 — —  Mixed Tocopherols G 111.4 — — L-Carnitine G26.30 3.285 3.28 Riboflavin G 3.181 1.166 1.4994 Tricalcium Phosphate G0-5230 12.5 41.89 Potassium Phosphate Monobasic G — 11.01 36.60 VitaminA Palmitate Mg — — 776.16  Vitamin A Palmitate Mg — — 427.19  AlphaTocopherol Mg — — 7.760 Potassium Phosphate Dibasic Kg 0-5.23  — — 1NKOH Kg as needed 1.583 as needed Potassium Hydroxide G as needed 79.15as needed

Control Formula 1 was prepared as described above in Examples 12-15;Control Formula 2 was prepared as described above in Examples 1-8, andControl Formula 3 was prepared as described above in Examples 9-11.

The buffering capacity and buffering strength of various days 1-2ready-to-feed (RTF) retort sterilized or reconstituted powder formulasand days 3-9 RTF retort sterilized, RTF aseptic sterilized, orreconstituted powder formulas was determined and compared to that ofControl Formulas 1-3 and to that of human milk. Specifically, thebuffering strength of the formulas (or human milk) was determined byadding 0.5 mL aliquots of 0.10 M HCl to 50 mL of each formula (orreconstituted formula, in the case of powder formula) at one minuteintervals. The pH of each formula was measured after each aliquotaddition. Buffering strength is reported as mL of 0.10 M HCl required tolower the pH of 50 mL of formula to 3.0. The buffering capacity of theformulas (or human milk) was determined by adding 5.00 mmoles of HCl to100 mL of each formula (or reconstituted formula, in the case of powderformula). The buffering capacity is reported as the increase in [H+]following the HCl addition. The results are shown in Table 6 below andin FIGS. 1 and 2.

TABLE 6 Energy Buffering Buffering Formula (kcal/L) Form Strength^(d)Capacity^(e) Control Formula 1 676 powder^(a) 25.8 0.776 mM Formula 14(days 3-9) 406 powder^(a) 17.1  9.55 mM Formula 14 (days 3-9)^(c) 406powder^(b) 17.0  9.33 mM Formula 12 (days 1-2) 270 powder^(b) 11.4  20.0mM Control Formula 2 676 retort 25.1 0.977 mM Formula 5 (days 3-9) 406retort 16.8  7.94 mM Formula 5 (days 3-9)^(c) 406 retort 16.2  9.12 mMFormula 2 (days 1-2) 270 retort 13.2  13.2 mM Formula 2 (days 1-2)^(c)270 retort 11.9  17.8 mM Formula 1 (days 1-2) 270 retort 10.8  18.6 mMControl Formula 3 676 aseptic 23.3  1.86 mM Formula 9 (days 3-9) 406aseptic 16.1  10.5 mM Human Milk 11.6  14.1 mM ^(a)Control Formula 1 wasreconstituted using 35.0 g of formula plus 240 mL of water prior todetermination of buffering capacity and buffering strength. ^(b)Formulas12 and 14 were reconstituted using 12.2 g of formula and 21.4 g offormula, respectively, plus 240 mL of water prior to determiningbuffering capacity and buffering strength. ^(c)Formulas 2, 5, and 14were tested twice. ^(d)as mL of 0.10M HCl required to lower the pH of 50mL of formula to 3.0. ^(e)as increase in [H+] upon addition of 5.00mmoles of HCl to 100 mL of formula.

As can be seen from these results, the buffering capacity of theformulations decreased with decreasing energy content. The days 1-2formulas, which had an energy content of 270 kcal/L, had the lowestbuffering capacity of all tested formulas. The buffering strength ofhuman milk has been reported to range from 9.0 to 18.0, with an averageof 13.5. As can be seen from the results set forth in Table 6 and FIGS.1 and 2, the buffering strength of the days 1-2 formulas was comparableto or lower than that of the tested human milk.

The decreased buffering capacity and buffering strength of the formulasof the present disclosure, and especially of the days 1-2 formulas, mayoffer physiological benefits to infants. In particular, decreasedbuffering capacity and strength may assist with achieving a morebeneficial gut microflora distribution, and may increase theeffectiveness of the inactivation of orally ingested intestinalpathogens.

Example 17

In this example, the effect of energy content on the buffering capacityand buffering strength of infant formula was evaluated. Specifically,the buffering capacity and buffering strength of days 1-2 (Formula 13)and days 3-9 (Formula 15) powder infant formulas of the presentdisclosure was determined following reconstitution and compared to thebuffering capacity and buffering strength of a commercially availablepowder control infant formula (Control Formula 1) followingreconstitution.

Formula 13 was reconstituted using 12.2 g of formula plus 240 mL ofwater, Formula 15 was reconstituted using 21.4 g of formula plus 240 mLof water, and Control Formula 1 was reconstituted using 35.0 g offormula plus 240 mL of water. The buffering capacity and bufferingstrength of each formula was determined. Specifically, the bufferingstrength of the formulas was determined by adding 1.00 mL aliquots of0.500 M HCl to 100 mL of reconstituted formula at one minute intervals.The pH of each formula was measured after each aliquot addition.Buffering strength is reported as mmoles of HCl required to lower the pHof 100 mL of the reconstituted formula from 6.00 to 3.00. The bufferingcapacity of the formulas was determined by adding 5.50 mmoles of HCl to100 mL of each reconstituted formula. The buffering capacity is reportedas the increase in [H+] following the HCl addition and the pH decreasefollowing the HCl addition. The results are shown in Table 7 below andin FIGS. 3-6.

TABLE 7 Formula 13 Formula 15 Control (days 1-2) (days 3-9) Formula 1Kcal/L 250 420 676 Buffering Strength 3.41 3.81 4.56 (mmoles) BufferingCapacity- 4.84 4.52 4.02 pH decrease Buffering Capacity- 6.17 mM 4.17 mM1.20 mM increase in [H+]

As can be seen from the results set forth in Table 7 and in FIGS. 3-6both the buffering strength and the buffering capacity (as measured byboth pH decrease and increase in [H+]) of the days 1-2 and days 3-9formulas were significantly lower than that of the control formula. Thedays 1-2 formula, which had an energy content of 250 kcal/L, had thelowest buffering capacity and buffering strength of all tested formulas,indicating that buffering strength and buffering capacity decreased withdecreasing energy content.

Example 18

In this example, the effect of energy content on the buffering capacityand buffering strength of infant formula was evaluated. Specifically,the buffering capacity and buffering strength of a 2 oz. retortsterilized days 1-2 infant formula of the present disclosure (Formula 3)was determined and compared to the buffering capacity and bufferingstrength of a 2 oz. commercially available retort sterilized controlinfant formula (Control Formula 2).

The buffering capacity and buffering strength of each formula wasdetermined. Specifically, the buffering strength of the formulas wasdetermined by adding 0.50 mL aliquots of 0.500 M HCl to 50 mL of eachformula at one minute intervals. The pH of each formula was measuredafter each aliquot addition. Buffering strength is reported as mmoles ofHCl required to lower the pH of 50 mL of the formula from 6.00 to 3.00.The buffering capacity of the formulas was determined by adding 2.75mmoles of HCl to 50 mL of each formula. The buffering capacity isreported as the increase in [H+] following the HCl addition and the pHdecrease following the HCl addition. The results are shown in Table 8below.

TABLE 8 Formula 3 Control (days 1-2) Formula 2 Kcal/L 250 676 BufferingStrength 1.53 2.28 (mmoles) Buffering Capacity- 4.34 4.13 pH decreaseBuffering Capacity- 10.7 mM 3.72 mM increase in [H+]

As can be seen from the results set forth in Table 8, both the bufferingstrength and the buffering capacity (as measured by both pH decrease andincrease in [H+]) of the days 1-2 formula were significantly lower thanthat of the control formula, indicating that buffering strength andbuffering capacity of the low calorie days 1-2 retort sterilized formulaof the present disclosure are lower than that of a conventional fullcalorie infant formula.

Example 19

In this example, the effect of energy content on the buffering capacityand the buffering strength of infant formula was evaluated.Specifically, the buffering capacity and buffering strength of a 32 oz.aseptic sterilized days 3-9 infant formula of the present disclosure(Formula 11) was determined and compared to the buffering capacity andbuffering strength of a 32 oz. commercially available aseptic sterilizedcontrol infant formula (Control Formula 3).

The buffering capacity and buffering strength of each formula wasdetermined. Specifically, the buffering strength of the formulas wasdetermined by adding 1.00 mL aliquots of 0.500 M HCl to 100 mL of eachformula at one minute intervals. The pH of each formula was measuredafter each aliquot addition. Buffering strength is reported as mmoles ofHCl required to lower the pH of 100 mL of the formula from 6.00 to 3.00.The buffering capacity of the formulas was determined by adding 5.50mmoles of HCl to 100 mL of each formula. The buffering capacity isreported as the increase in [H+] following the HCl addition and the pHdecrease following the HCl addition. The results are shown in Table 9below.

TABLE 9 Formula 11 Control (days 3-9) Formula 3 Kcal/L 410 676 BufferingStrength 3.46 3.84 (mmoles) Buffering Capacity- 4.78 4.54 pH decreaseBuffering Capacity- 8.51 mM 5.50 mM increase in [H+]

As can be seen from the results set forth in Table 9, both the bufferingstrength and the buffering capacity (as measured by both pH decrease andincrease in [H+]) of the days 3-9 formula were significantly lower thanthat of the control formula, indicating that buffering strength andbuffering capacity of the low calorie days 3-9 aseptic sterilizedformula of the present disclosure is lower than that of a conventionalfull calorie infant formula.

Example 20

In this example, the effect of the energy content of infant formula onthe rate and extent of protein hydrolysis was evaluated. Specifically,the extent of protein hydrolysis of reconstituted days 1-2 (Formula 13)and reconstituted days 3-9 (Formula 15) powdered infant formulas of thepresent disclosure was determined following an in vitro gastrointestinaldigestion, and compared to the extent of protein hydrolysis of areconstituted powder control infant formula (Control Formula 1).

Formula 13 was reconstituted using 12.2 g of formula plus 240 mL ofwater, Formula 15 was reconstituted using 21.4 g of formula plus 240 mLof water, and Control Formula 1 was reconstituted using 35.0 g offormula plus 240 mL of water. Digests were prepared by subjecting thereconstituted formulas to an in vitro gastrointestinal digestion.Specifically, the pH of 40 mL of each reconstituted formula was adjustedto 4.5 using 6 M HCl. 1.00 mL of USP pepsin, prepared in 56 mg/mL ofwater, was added to the formula, and the resulting mixture was stirredat room temperature for one hour. The pH of the mixture was thenadjusted to 7.2 using 10 N NaOH. 4.00 mL of USP pancreatinamylase/protease, prepared in 6.94 mg/mL water, plus USP pancreatinlipase, prepared in 6.94 mg/mL water, was then added, and the mixturewas stirred at room temperature for two hours. The resulting digestswere centrifuged at 31,000×g at 20° C. for 4 hours.

The supernatant was analyzed by HPLC using a Superdex® Peptide 10/300 GLgel filtration column (Amersham Biosciences). Specifically, 5 mg of thesupernatant was added to 1 mL of a mobile phase solution (700 mLMilli-Q® water, 300 mL acetonitrile, 1.00 mL TFA) and the resultingsolution was run at ambient temperature on the Superdex® column (flowrate: 0.4 mL/minute; detection: UV at 205 nm; injection: 10 μL; runtime: 80 minutes) to determine the molecular weight median of theprotein in the digests and the amount of protein having a molecularweight of greater than 5000 Daltons, as a percentage of total protein,in the digests. These determinations are indicators of the extent ofprotein digestion. The pellet produced following centrifugation of thedigests was also tested for the presence of insoluble protein using acidhydrolysis/amino acid profile using conventional methods. The resultsare shown in Table 10 below and in FIGS. 7-9.

TABLE 10 Formula 13 Formula 15 Control (days 1-2) (days 3-9) Formula 1Kcal/L 250 420 676 Protein MW median (Da) 777 925 1022 Protein >5000 Da8.4% 13.4% 14.0% (% total protein) Insoluble protein^(a) (mg/L) 24 59156 ^(a)total protein in the pellet after high speed centrifugation ofthe digest

As can be seen from these results, the protein hydrolysis was moreextensive in the days 1-2 and days 3-9 formulas than in the controlformula. Further, all three digestion indicators (protein MW median,amount of protein >5000 Da, and amount of insoluble protein) decreasedwith decreasing energy content. These results indicate that the rate ofprotein digestion is inversely correlated with energy content.

Example 21

In this example, the effect of the energy content of infant formula onthe rate and extent of protein hydrolysis was evaluated. Specifically,the extent of protein hydrolysis of a 2 oz. retort sterilized days 1-2infant formula of the present disclosure (Formula 3) was determinedfollowing an in vitro gastrointestinal digestion, and compared to theextent of protein hydrolysis of a 2 oz. commercially available retortsterilized control infant formula (Control Formula 2).

Digests were prepared by subjecting the formulas to an in vitrogastrointestinal digestion using the procedure set forth in Example 20.The digests were centrifuged at 31,000×g at 20° C. for 4 hours. Thesupernatant was analyzed by HPLC using a Superdex® Peptide 10/300 GL gelfiltration column (Amersham Biosciences) using the procedure set forthabove in Example 20, and the molecular weight median of the protein inthe digests and the amount of protein having a molecular weight ofgreater than 5000 Daltons, as a percentage of total protein, in thedigests was determined. The pellet produced following centrifugation ofthe digests was also tested for the presence of insoluble protein usingthe acid hydrolysis/amino acid profile technique described in Example20. The results are shown in Table 11 below.

The digests were also tested for the presence of the Maillard reactionmarker furosine using acid hydrolysis and HPLC. These results are alsoshown in Table 11 below.

TABLE 11 Formula 3 Control (days 1-2) Formula 2 Kcal/L 250 676 ProteinMW median (Da) 789 992 Protein >5000 Da (% total protein) 3.77% 8.81%Insoluble protein^(a) (mg/L) 48 471 Furosine (mole % of total lysine)0.84% 2.61% ^(a)total protein in the pellet after high speedcentrifugation of the digest

As can be seen from these results, the protein hydrolysis was moreextensive in the days 1-2 formula than in the control formula. All threedigestion indicators (protein MW median, amount of protein >5000 Da, andamount of insoluble protein) decreased with decreasing energy content.These results indicate that the rate of protein digestion is inverselycorrelated with energy content. Further, the days 1-2 formula had lowerlevels of the Maillard reaction marker furosine than did the controlformula. These results suggest that the low calorie days 1-2 retortsterilized formulas of the present disclosure are less susceptible toMaillard reactions than conventional full calorie infant formulas.

Example 22

In this example, the effect of the energy content of infant formula onthe rate and extent of protein hydrolysis was evaluated. Specifically,the extent of protein hydrolysis of a 32 oz. aseptic sterilized days 3-9infant formula of the present disclosure (Formula 11) was determinedfollowing an in vitro gastrointestinal digestion, and compared to theextent of protein hydrolysis of a 32 oz. commercially available asepticsterilized control infant formula (Control Formula 3).

Digests were prepared by subjecting the formulas to an in vitrogastrointestinal digestion using the procedure set forth in Example 20.The digests were centrifuged at 31,000×g at 20° C. for 4 hours. Thesupernatant was analyzed by HPLC using a Superdex® Peptide 10/300 GL gelfiltration column (Amersham Biosciences) using the procedure set forthabove in Example 20, and the molecular weight (MW) median of the proteinin the digests and the amount of protein having a molecular weight ofgreater than 5000 Daltons, as a percentage of total protein, in thedigests was determined. The pellet produced following centrifugation ofthe digests was also tested for the presence of insoluble protein usingthe acid hydrolysis/amino acid profile technique described in Example20. The results are shown in Table 12 below.

TABLE 12 Formula 11 Control (days 3-9) Formula 3 Kcal/L 410 676 ProteinMW median (Da) 799 978 Protein >5000 Da (% total protein) 2.5% 9.5%Insoluble protein^(a) (mg/L) 110 400 ^(a)total protein in the pelletafter high speed centrifugation of the digest

As can be seen from these results, the protein hydrolysis was moreextensive in the days 3-9 formula than in the control formula. All threedigestion indicators (protein MW median, amount of protein >5000 Da, andamount of insoluble protein) decreased with decreasing energy content.These results indicate that the rate of protein digestion is inverselycorrelated with energy content.

Example 23

In this example, the effect of the energy content of infant formula onthe rate and extent of protein hydrolysis was evaluated. Specifically,the extent of protein hydrolysis of reconstituted days 1-2 (Formula 13)and reconstituted days 3-9 (Formula 15) powdered infant formulas of thepresent disclosure was determined following digestion with pancreatin,and compared to the extent of protein hydrolysis of a reconstitutedcommercially available powder control infant formula (Control Formula 1)following pancreatin digestion.

Formula 13 was reconstituted using 12.2 g of formula plus 240 mL ofwater, Formula 15 was reconstituted using 21.4 g of formula plus 240 mLof water, and Control Formula 1 was reconstituted using 35.0 g offormula plus 240 mL of water. Digests were prepared by subjecting thereconstituted formulas to digestion with pancreatin. Specifically, 9.00mL of 0.05 M NaH₂PO₄ (pH 7.5) was added to 9.00 mL of each formula in a20 mL vial. 2.00 mL of porcine pancreatin, prepared at 4.0 g/L in pH 7.5buffer, was added to the formula, and the vial was placed in a 37° C.water bath for 71 minutes. After 71 minutes, a 1.5 mL aliquot of themixture was transferred into an HPLC autosampler vial, and the vial wascrimp sealed. The sealed vial was placed in a 100° C. heating module for5 minutes to terminate the pancreatin digestion. 0.400 mL of theresulting digest was diluted with 1.00 mL of 8.30/6.00/0.02 (v/v) ofwater/acetonitrile/trifluoroacetic acid. The diluted digest wascentrifuged at 14,000×g at room temperature for 5 minutes. Thesupernatant was analyzed by HPLC using a Superdex® Peptide 10/300 GL gelfiltration column (Amersham Biosciences) using the procedure set forthabove in Example 20, and the molecular weight (MW) median of the proteinin the digests and the amount of protein having a molecular weight ofgreater than 5000 Daltons, as a percentage of total protein, in thedigests was determined. The results are shown in Table 13 below and inFIGS. 10 and 11.

TABLE 13 Formula 13 Formula 15 Control (days 1-2) (days 3-9) Formula 1Kcal/L 250 420 676 Protein MW median (Da) 680 748 853 Protein >5000 Da(% total 2.15% 2.54% 3.03% protein)

As can be seen from these results, the protein hydrolysis was moreextensive in the days 1-2 and days 3-9 formulas than in the controlformula. Further, both digestion indicators (protein MW median, amountof protein >5000 Da) decreased with decreasing energy content. Theseresults indicate that the rate of protein digestion is inverselycorrelated with energy content.

Example 24

In this example, the effect of the energy content of infant formula onthe rate and extent of protein hydrolysis was evaluated. Specifically,the extent of protein hydrolysis of a 2 oz. retort sterilized days 1-2infant formula of the present disclosure (Formula 3) was determinedbefore and after pancreatin digestion, and compared to the extent ofprotein hydrolysis of a 2 oz. commercially available retort sterilizedcontrol infant formula (Control Formula 2) before and after pancreatindigestion.

Digests were prepared by subjecting the formulas to pancreatin digestionusing the same procedure as set forth in Example 23, except the infantformula/pancreatin mixture was held in the 37° C. water bath for only 60minutes. The diluted digests were centrifuged at 14,000×g at roomtemperature for 5 minutes. The supernatant as well as a sample of theinfant formulas prior to digestion were analyzed by HPLC using aSuperdex® Peptide 10/300 GL gel filtration column (Amersham Biosciences)using the procedure set forth above in Example 20, and the molecularweight median of the protein in the infant formula prior to digestionand the molecular weight median of the protein following 60 minutes ofpancreatin digestion was determined. The results are shown in Table 14below.

TABLE 14 Formula 3 Control (days 1-2) Formula 2 Kcal/L 250 676 ProteinMW median 14,774 19,120 (Da) before digestion Protein MW median 801 1128(Da) after 60 min. digestion

As can be seen from these results, the rate of protein hydrolysis wasfaster in the low calorie days 1-2 formula than in the control formula.Further, the MW median values at 60 minutes of pancreatin digestion wereproportional to the caloric density of the infant formulas, indicatingthat protein digestion rate was inversely correlated to energy content.

Example 25

In this example, the effect of the energy content of infant formulas onthe rate and extent of protein hydrolysis was evaluated. Specifically,the extent of protein hydrolysis of reconstituted days 1-2 (Formula 12)or days 3-9 (Formula 14) powdered infant formulas, days 1-2 (Formulas 1and 2) or days 3-9 (Formula 5) 2 oz. retort sterilized infant formula,and days 3-9 (Formula 9) 32 oz. aseptic sterilized infant formula of thepresent disclosure was determined following pancreatin digestion(powders) or in vitro GI digestion (liquids) and compared to the extentof protein hydrolysis of a reconstituted commercially available powdercontrol infant formula (Control Formula 1), a 2 oz. commerciallyavailable retort sterilized control infant formula (Control Formula 2),and a 32 oz. commercially available aseptic sterilized control formula(Control Formula 3).

Formula 12 was reconstituted using 12.2 g of formula plus 240 mL ofwater, Formula 14 was reconstituted using 21.4 g of formula plus 240 mLof water, and Control Formula 1 was reconstituted using 35.0 g offormula plus 240 mL of water. Digests were prepared by subjecting theformulas (or reconstituted formulas) to pancreatin digestion using thesame procedure as set forth above. The supernatant was analyzed by HPLCusing a Superdex® Peptide 10/300 GL gel filtration column (AmershamBiosciences) using the procedure set forth above in Example 20, and themolecular weight (MW) median of the protein in the digests and theamount of protein having a molecular weight of greater than 5000Daltons, as a percentage of total protein, in the digests wasdetermined. The results are shown in Table 15 below.

TABLE 15 Protein MW Energy >5000 Da (% Protein MW Formula (kcal/L) Formtotal protein) median (Da) Control Formula 1 676 powder 17.9% 1050Formula 14 (days 3-9)^(a) 406 powder 10.9% 846 Formula 14 (days 3-9) 406powder 8.4% 812 Formula 12 (days 1-2) 270 powder 5.2% 717 ControlFormula 2 676 retort 13.7% 988 Formula 5 (days 3-9) 406 retort 5.3% 789Formula 1 (days 1-2) 270 retort 3.9% 730 Formula 2 (days 1-2) 270 retort2.9% 707 Control Formula 3 676 aseptic 10.2% 963 Formula 9 (days 3-9)406 aseptic 4.1% 801 ^(a)Formula 14 was tested twice.

As can be seen from these results, the protein hydrolysis was moreextensive in the days 1-2 and days 3-9 formulas than in the controlformulas. Further, both digestion indicators (protein MW median, amountof protein >5000 Da) decreased with decreasing energy content. Theseresults indicate that the rate of protein digestion is inverselycorrelated with energy content.

Example 26

In this example, the effect of micronutrient content on the emulsionstability of days 1-2 retort sterilized infant formula and on days 3-9aseptic sterilized infant formula was evaluated. Specifically, theemulsion stability of 32 oz. days 3-9 aseptic sterilized infant formulashaving either a high (Formula 11) or low (Formula 9) micronutrientcontent and 2 oz. days 1-2 retort sterilized infant formulas havingeither a high (Formula 3) or low (Formula 1) micronutrient content wascompared.

Protein loading levels, expressed as the protein percent of the creamlayer formed following high speed centrifugation of the formula, wereused to determine emulsion stability. Protein loading levels for eachformula were determined by pouring 36-38 grams of formula into a tared50 mL centrifugation tube, and capping the tubes. The capped tubes werethen placed in a JA-20 fixed angle rotor (Beckman Coulter, P/N 334831),and the rotor was placed into a Beckman J2-HS centrifuge (BeckmanCoulter). The samples were centrifuged at 31,000×g at 20° C. for 8hours. Following centrifugation, a cream layer formed on the sample. Thecream layer was transferred into a tared beaker, and its weightrecorded. The supernatant was poured into a separate beaker, and thetube was reweighed to determine the weight of the pellet.

The amount of protein in the cream layer was determined using an acidhydrolysis/amino acid determination technique. The results are set forthin Table 16 below.

TABLE 16 Protein % of Micro- cream layer Energy nutrient (approximate %Formula (kcal/L) content Form w/w) Formula 11 (days 3-9) 410 highaseptic 5.1% Formula 9 (days 3-9) 406 low aseptic 4.7% Formula 3 (days1-2) 250 high retort 4.6% Formula 1 (days 1-2) 270 low retort 5.9%Average (n = 4) 5.1% ± 0.6%

Protein loading values are indicators of emulsion stability.Specifically, emulsion stability generally increases with increasingprotein loading values. As can be seen from the above-results, theprotein loading values were higher in the days 1-2 retort sterilizedformula having a low micronutrient content (i.e., Formula 1) than in thedays 1-2 retort sterilized formula having a high micronutrient content(i.e., Formula 3). These results indicate that there is increasedemulsion stability in days 1-2 retort sterilized formulas having lowmicronutrient content, as compared to comparable formulas having highmicronutrient content. No significant difference in protein loading wasseen between the high micronutrient content and low micronutrientcontent aseptic sterilized formulas.

Example 27

In this example, the effect of micronutrient content on the emulsionstability of days 3-9 retort sterilized formulas was evaluated.Specifically, the emulsion stability of 2 oz. days 3-9 retort sterilizedinfant formulas having either a high (Formula 8) or low (Formula 6)micronutrient content was compared.

Protein loading levels, expressed as the protein percent of the creamlayer formed following high speed centrifugation of the formula, wereused to determine emulsion stability. Protein loading levels for eachformula were determined using the procedure set forth in Example 26. Theamount of cream layer, by weight of the whole product, and the amount ofproteins in the cream layer, by weight of the whole product, were alsocalculated. The results are set forth in Table 17 below.

TABLE 17 Cream layer Protein % of protein % of Energy Micronutrientcream layer whole product Formula (kcal/L) content (w/w) (w/w) Formula 6406 low 6.9% 0.35% (days 3-9) Formula 8 410 high 5.1% 0.22% (days 3-9)

As can be seen from these results, the protein loading values werehigher in Formula 6, which had a low micronutrient content, than in thehigh micronutrient formula (I.e., Formula 8). Formula 6 also formed alarger cream layer, and had a higher percentage of proteins in the creamlayer, by weight of the whole product, than did Formula 8. These resultsindicate that there is increased emulsion stability in days 3-9 retortsterilized formulas having a low micronutrient content, as compared tocomparable formulas having a high micronutrient content. The lowmicronutrient content days 3-9 retort sterilized formula (I.e., Formula6) also had a higher protein loading value, and thus an increasedemulsion stability, as compared to low micronutrient content days 1-2retort sterilized formulas (see Formula 1, Example 26).

Example 28

In this example, the effect of micronutrient content on the color ofdays 1-2 and days 3-9 retort sterilized formulas and on days 3-9 asepticsterilized formulas was evaluated.

Color quality of the formulas was evaluated using the Agtron colormethod. The Agtron color method measures the percent of light reflectedfrom the sample on a scale of 0 (black) to 100 (white) using aspectrophotometer. Brighter colored infant formulas, which are typicallypreferred by consumers, have a higher Agtron color score, while darkercolored formulas have a lower score. The Agtron color scores for low andhigh micronutrient content retort and aseptic formulas of the presentdisclosure, measured at various time periods, are set forth in Table 18(retort formulas) and Table 19 (days 3-9 aseptic formulas) below.

TABLE 18 Retort Formulas Energy Micronutrient Time Agtron color Formula(kcal/L) content interval score (%)^(a) Formula 3 250 high 0 days 39.3(days 1-2)  1 mo. —  2 mo. 33.3  4 mo. 30.2  9 mo. 28.5 12 mo. 28.2Formula 4 250 high 0 days 44.1 (days 1-2)  1 mo. —  3 mo. 37.5  6 mo.35.4  9 mo. 33.4 12 mo. 33.0 Formula 1 270 low 0 days 47.9 (days 1-2)  2mo. 43.7  4 mo. 42.2  6 mo. 40.3  9 mo. 38.6 Formula 2 270 low 0 days54.4 (days 1-2)  3 mo. 49.7  6 mo. 47.8 Formula 8 410 high 0 days 39.4(days 3-9) Formula 5 406 low 0 days 51.1 (days 3-9)  3 mo. 48.8  6 mo.46.0 Formula 6 406 low 0 days 45.3 (days 3-9) Formula 7 406 low 0 days46.2 (days 3-9) (—) means not tested ^(a)Agtron color scores weredetermined using an Agtron M-45 spectrophotometer (blue filter - 436 nm)for all measurements.

TABLE 19 Days 3-9 Aseptic Formulas Energy Micronutrient Time Agtroncolor Formula (kcal/L) content interval score (%)^(a) Formula 11 410high 0 days 53.1  1 mo. 49.7  2 mo. —  4 mo. — 12 mo. 46.2 Formula 10410 high 0 days 56.5  1 mo. —  3 mo. 51.7  6 mo. 53.1  9 mo. 51.4 12 mo.47.6 Formula 9  406 low 0 days 61.5  1 mo. —  2 mo. 60.0  6 mo. 56.9  9mo. 53.8 (—) means not tested ^(a)Agtron color scores were determinedusing an Agtron M-45 spectrophotometer (blue filter - 436 nm) for allmeasurements.

As can be seen from these results, the retort sterilized days 1-2 infantformulas having a low micronutrient content had a higher Agtron colorscore, and thus a brighter colored appearance, than retort sterilizeddays 1-2 infant formulas having a high micronutrient content. Similarresults were obtained for the days 3-9 retort formulas and the days 3-9aseptic formulas, where the low micronutrient content formulas had ahigher Agtron color score than comparable formulas having a highmicronutrient content. The improved color of the low micronutrientformulas, as compared to comparable high micronutrient formulas, wasalso observed even after extended periods of time, in some cases up to 9months following product formulation. These results indicate that infantformulas of the present disclosure that have a low micronutrient contenthave a brighter and lighter colored appearance than comparable formulasthat have a high micronutrient content.

Example 29

In this example, the effect of micronutrient content on the particlesize distribution and creaming velocity of retort sterilized days 1-2formulas was evaluated.

Specifically, the particle size distribution of 2 oz. retort sterilizeddays 1-2 formulas having either a high micronutrient content (Formula 3)or a low micronutrient content (Formula 1) was determined using aBeckman Coulter LS 13 320 light scattering machine. The results areshown in FIG. 12.

As can be seen from FIG. 12, the majority of the particles in the lowmicronutrient days 1-2 retort formula (Formula 1) were between about 0.1μm and about 0.8 μm in size, with a smaller number of particles rangingfrom about 1 μm to about 8 μm. In contrast, the particle sizedistribution of the high micronutrient days 1-2 retort formula (Formula3) ranged more equally from about 0.1 μm to about 7 μm.

The average particle size for each formula was determined from theparticle size distribution and was used to calculate the creamingvelocity of each formula. Specifically, the creaming velocity wascalculated using the following equation:

$v_{cream} = {\frac{2}{9}\frac{\rho_{fluid} - \rho_{particle}}{\eta}{gR}^{2}}$

wherein:v_(cream) is the creaming velocityρ_(fluid) is the density of the formulaρ_(particle) is the density of the particlesη is the viscosity of the formulaR is the average particle sizeg is the gravitational acceleration

The density of the particles (e.g., oil droplets) was calculated bymeasuring the total surface area of the particles in a unit sample (100mL) using a Beckman Coulter LS 13 320 light scattering machine. Thevolume of protein attached to the surface of the oil droplets was thenmeasured using ultracentrifugation. The protein volume was then dividedby the total surface area of the oil droplets to get the averagethickness of the protein layer coated on each oil droplet. The averageparticle density was then calculated using 1.41 for the density ofprotein (Fischer, et al., Protein Science (2004), Vol. 13 (10), p.2825-2828).

R² values and the creaming velocity for each formula are shown in Table20.

TABLE 20 Particle Size and Creaming Velocity of Days 1-2 Retort FormulasSquare of Creaming Energy Micronutrient average particle velocity(kcal/L) content size (R²) (μm²) (cm/day) Formula 1 270 low 1.8 3.2Formula 3 250 high 3.5 6.3

As can be seen from this table, the average particle size of the lowmicronutrient days 1-2 retort formula (Formula 1) was smaller than thatof the high micronutrient days 1-2 retort formula (Formula 3). Since asmaller particle size may be representative of product stability, theseresults suggest that the low micronutrient days 1-2 retort formulas ofthe present disclosure have a greater product stability than comparableformulas having a high micronutrient content.

Creaming velocity measures the rate of movement of particles (e.g.,droplets) through a liquid sample, in this instance, the infant formula,and is predictive of the capacity of the infant formula to form a creamlayer. As can be seen from Table 20, the creaming velocity of the lowmicronutrient content days 1-2 retort formula was lower than that of thehigh micronutrient content days 1-2 retort formula. These resultsindicate that the low micronutrient content days 1-2 retort formulas ofthe present disclosure have a reduced capacity to form a cream layer,and thus have improved physical stability as compared to comparable highmicronutrient formulas.

1-25. (canceled)
 26. A low micronutrient infant formula comprisingmicronutrients and at least one macronutrient selected from the groupconsisting of protein, carbohydrate, fat, and combinations thereof, andhaving an energy content of 200 to less than 600 kilocalories per literof formula, where the micronutrients include at least a combination ofcopper, phosphorous, iron, calcium, and zinc, at least two of which arepresent in an amount of at least 20% less on a per volume basis than thefollowing conventional amounts: 728 μg/L copper when in an infantformula in the form of an aseptic liquid, 676 μg/L copper when in aninfant formula in the form of a retort liquid, 720 μg/L copper when inan infant formula in the form of a reconstituted powder; 341 mg/Lphosphorous when in an infant formula in the form of an aseptic liquid,349 mg/L phosphorous when in an infant formula in the form of a retortliquid, 332 mg/L phosphorous when in an infant formula in the form of areconstituted powder; 13.7 mg/L iron when in an infant formula in theform of an aseptic liquid, 13.4 mg/L iron when in an infant formula inthe form of a retort liquid, 13.9 mg/L iron when in an infant formula inthe form of a reconstituted powder; 581 mg/L calcium when in an infantformula in the form of an aseptic liquid, 585 mg/L calcium when in aninfant formula in the form of a retort liquid, 580 mg/L calcium when inan infant formula in the form of a reconstituted powder; 6.67 mg/L zincwhen in an infant formula in the form of an aseptic liquid, 6.46 mg/Lzinc when in an infant formula in the form of a retort liquid, and 6.69mg/L zinc when in an infant formula in the form of a reconstitutedpowder.
 27. An infant formula according to claim 26, wherein the formulais a retort sterilized liquid, an aseptically sterilized liquid, aconcentrated liquid, or a powder.
 28. An infant formula according toclaim 26, wherein the energy content of the infant formula is 250 to 500kilocalories per liter of formula.
 29. An infant formula according toclaim 26, comprising from 0.5 g to 1.0 g protein per 100 mL of formula,1.2 g to 2.5 g fat per 100 mL of formula, and 2.7 g to 6.5 gcarbohydrate per 100 mL of formula.
 30. An infant formula according toclaim 26, wherein the infant formula has a protein loading level of atleast 5%.
 31. An infant formula according to claim 26, wherein theinfant formula has a creaming velocity of from 1 cm/day to 5 cm/day. 32.An infant formula according to claim 26, wherein the energy content ofthe infant formula is about 270 kilocalories per liter of formula. 33.An infant formula according to claim 26, wherein the infant formula hasan energy content of 200 to 360 kilocalories per liter of formula. 34.An infant formula according to claim 33, wherein the infant formula is adays 1-2 infant formula.
 35. An infant formula according to claim 26,wherein the infant formula has an energy content of 360 to 600kilocalories per liter of formula.
 36. An infant formula according toclaim 35, comprising 0.76 g to 1.0 g protein per 100 mL of formula, 1.8g to 2.5 g fat per 100 mL of formula, and 4.1 g to 6.5 g carbohydrateper 100 mL of formula.
 37. An infant formula according to claim 35,wherein the infant formula is a days 3-9 infant formula.
 38. An infantformula according to claim 26, wherein the infant formula has abuffering capacity expressed as the H+ concentration following additionof 5 mmoles of HCl to 100 mL of formula of 2 mM to 25 mM.
 39. An infantformula according to claim 26, wherein the infant formula has abuffering strength expressed as the mL of 0.1 M HCl needed to decreasethe pH of 50 mL of formula from a starting pH of 6.0 to a pH of 3.0 of 9mL to 18 mL.
 40. An infant formula according to claim 26, wherein theinfant formula contains micronutrients in an amount that is low enoughto provide the infant formula with an Agtron color score two monthsafter formulation of at least
 40. 41. A kit comprising: at least onedays 1-2 low micronutrient infant formula having an energy content of200 to 360 kilocalories per liter of formula; and at least one days 3-9low micronutrient infant formula having an energy content of 360 to 600kilocalories per liter of formula.
 42. A method for providing nutritionto an infant, the method comprising administering to the infant a lowmicronutrient infant formula according to claim 26.